Polycomb group genes play crucial roles in the maintenance of the transcriptionally silenced state of genes for proper cell differentiation in animals and plants. While components of the polycomb repressive complex2 ...Polycomb group genes play crucial roles in the maintenance of the transcriptionally silenced state of genes for proper cell differentiation in animals and plants. While components of the polycomb repressive complex2 （PRC2） are evolutionarily conserved and their functions are extensively studied in plants, PRCI differs considerably between animals and plants, and its functions in plants are as yet not well described. Previous studies have identified the Arabidopsis AtRINGla and AtRINGlb as homologues of the animal PRC1 subunit RING1. Here, we show that the Atringla Atringlb double mutant exhibits derepression of embryonic traits during vegetative growth. Accordingly, several key regulatory genes involved in embryogenesis and stem cell activity are ectopically expressed in the mutant. Furthermore, we show that the mutant phenotypes and increased expression of regulatory genes are enhanced by the PRC2 mutant c/f. Finally, we show that three homologues of the animal PRCl-subunit ring-finger protein BMI1, AtBMIIa, AtBMIlb and AtBMIlc, can bind with AtRINGla or AtRINGIb, and in addition, AtBMIlc can bind with LHP1. The Atbmila Atbmilb double mutant shows derepression of embryonic traits similar to that of the Atringla Atringlb double mutant. Interestingly, expression levels of AtBMIla, AtBMIlb and AtBMIlc are elevated in the Atringla Atringlb mutant and those of AtBMIlc, AtRINGla and AtRINGlb are elevated in the Atbmila Atbmilb mu- tant, suggesting a self-regulatory feedback mechanism. Taken together, our results illuminate crucial functions of the PRCl-like ring-finger components in stable repression of embryonic traits and regulatory genes for proper somatic growth.展开更多
Endocytosis occurs at the cell surface and involves internalization of the plasma membrane （PM） along with its constituent membrane proteins and lipids. Endocytosis is involved in sampling of the extracellular milie...Endocytosis occurs at the cell surface and involves internalization of the plasma membrane （PM） along with its constituent membrane proteins and lipids. Endocytosis is involved in sampling of the extracellular milieu and also serves to regulate various processes initiated at the cell surface. These include nutrient uptake, signaling from cell- surface receptors, and many other processes essential for cell and tissue functioning in metazoans. It is also central to the maintenance of PM lipid and protein homeostasis. There are multiple means of internalization that operate concurrently, at the cell surface. With advancement in high-resolution visualization techniques, it is now possible to track multiple endocytic cargo at the same time, revealing a remarkable diversity of endocytic processes in a single cell. A combination of live cell imaging and efficient genetic manipulations has also aided in understanding the functional hierarchy of molecular players in these mechanisms of internalization. Here we provide an account of various endocytic routes, their mechanisms of operation and occurrence across phyla.展开更多
Drosophila RYBP （dRYBP; Ringl and Y_Y1 Binding Protein） is a Polycomb and trithorax interacting protein, whose homologous RYBP/DEDAF mammalian counterparts exhibit tumor cell-specific killing activity. Here we show ...Drosophila RYBP （dRYBP; Ringl and Y_Y1 Binding Protein） is a Polycomb and trithorax interacting protein, whose homologous RYBP/DEDAF mammalian counterparts exhibit tumor cell-specific killing activity. Here we show that although endogenous dRYBP is not involved in developmental apoptosis, high levels of exogenous dRYBP induce apoptosis in all the imaginal discs of the fly, indicating that dRYBP apoptotic activity is not specific to tumor cells. We also show that dRYBP-induced apoptosis is inhibited by high levels of either p35 or DIAP1 （Drosophila Inhibitor of Apoptosis Protein 1）, and requires the function of the pro-apoptotic REAPER, HID and GRIM proteins, the apical caspase DREDD, the adaptor dFADD protein as well as TRITHORAX （TRX）, an epigenetic transcriptional regulator. Furthermore, we demonstrate that overexpression of TRX also induces apoptosis in the imaginal discs. Finally, we show that the expression of reaper-lacZ is upregulated both upon dRYBP-induced apoptosis and upon TRX- induced apoptosis in imaginal discs and that the reaper gene is a direct target of dRYBP in Drosophila embryos. Our results indicate that dRYBP triggers in a receptor-mediated apoptotic pathway that also includes TRX-dependent epigenetic regulation of gene expression.展开更多
While severe acute respiratory syndrome coronavirus （SARS-CoV）~as initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that yirus entry may also involve endo...While severe acute respiratory syndrome coronavirus （SARS-CoV）~as initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that yirus entry may also involve endocytosis. We have found that SARS-CoV enters cells viapH- and receptor-dependent endocytosis. Treatment of cells with either SARS-COV spike protein or spike-bearing pseudoviruses resulted in the translocation of angiotensin-converting enzyme 2 （ACE2）, the functional receptor of SARS-CoV, from the cell surface to endosomes. In addition, the spike-bearing pseudoviruses and early endosome antigen 1 were found to colocalize in endosomes. Further analyses using specific endocytic path- way inhibitors and dominant-negative Epsl5 as well as caveolin-1 colocalization study suggested that virus entry was mediated by a clathrin- and caveolae-independent mechanism. Moreover, cholesterol- and sphingolipid-rich lipid raft microdomains in the plasma membrane, which have been shown to act as platforms for many physiological signaling pathways, were shown to be involved in virus entry. Endocytic entry of SARS-CoV may expand the cellular range of SARS-CoV infection, and our findings here contribute to the understanding of SARS-CoV pathogenesis, providing new information for anti-viral drug research.展开更多
The transcription factor p53 is a key regulator of the DNA damage response to genotoxic stress in higher eukaryotes. Mutation/inactivation of p53 has been implicated in the pathogenesis of many tumor types [ 1 ]. p53 ...The transcription factor p53 is a key regulator of the DNA damage response to genotoxic stress in higher eukaryotes. Mutation/inactivation of p53 has been implicated in the pathogenesis of many tumor types [ 1 ]. p53 responds to cellular insults, such as DNA damage or hypoxia, either by arresting the cell cycle so that DNA damage could be repaired, or by triggering apoptosis ifDNA repair is futile. Transcriptional activation of apoptosis-related genes by p53 is critical for the induction of programmed cell death. In search of target genes of p53 involved in cell-cycle arrest and/or apoptosis, numerous p53-inducible genes have been identified over the past 15 years. Among them are genes such as Bax [2], Puma [3], Noxa [4], Bid [5], p21waf1 [6], and many others. These apoptosis-related factors function in different cell organelles including the cytosolic membrane, the mitochondria and the cytosol. However, none of these candidate effectors alone can be accountable for the complicated mechanisms underlying the p53 transcription-dependent apoptotic signaling pathways. Earlier work implicated that Bad may contribute to p53-induced mitochondria-mediated apoptosis, yet no known molecular mechanism had been ascribed to this observation. Our recent study published in Molecular and Cellular Biology revealed that a functional p53-binding element exists at the human bad genomic locus, residing roughly 6.6 kb upstream of the Bad translation start codon [7]. The fact that this p53-binding element is relatively far away from the Bad promoter could be the reason why it has not been recognized as a p53-binding element until today. Interestingly enough, we found that a similar p53-binding region is well conserved approximately 7 kb upstream of the murine bad locus （our unpublished data）.展开更多
The application of genomic technologies to the study of mRNA processing is increasingly conducted in metazoan organisms in order to understand the complex events that occur during and after transcription. Large-scale ...The application of genomic technologies to the study of mRNA processing is increasingly conducted in metazoan organisms in order to understand the complex events that occur during and after transcription. Large-scale systems analyses ofmRNA-protein interactions and mRNA dynamics have revealed specificity in mRNA transcription, splicing, transport, translation, and turnover, and have begun to make connections between the different layers ofmRNA processing. Here, we review global studies of post-transcriptional processes and discuss the challenges facing our understanding of mRNA regulation in metazoan organisms. In parallel, we examine genome-scale investigations that have expanded our knowledge of RNA-binding proteins and the networks of mRNAs that they regulate.展开更多
Thioredoxin-1 （TRX） is a stress-inducible redox-regulatory protein with antioxidative and anti-inflammatory effects. Here we show that the release of histamine from mast cells elicited by cross-linking of high-affin...Thioredoxin-1 （TRX） is a stress-inducible redox-regulatory protein with antioxidative and anti-inflammatory effects. Here we show that the release of histamine from mast cells elicited by cross-linking of high-affinity receptor for IgE （FcεRI） was significantly suppressed in TRX transgenic （TRX-tg） mice compared to wild type （WT） mice. Intracellular reactive oxygen species （ROS） of mast cells stimulated by IgE and antigen was also reduced in TRX-tg mice compared to WT mice. Whereas there was no difference in the production ofcytokines （IL-6 and TNF-α） from mast cells in response to 2,4-dinitrophenylated bovine serum albumin （DNP-BSA） stimulation in TRX-tg and WT mice. Immunological status of TRX-tg mice inclined to T helper （Th） 2 dominant in primary immune response, although there was no difference in the population of dendritic cells （DCs） and regulatory T cells. We conclude that the histamine release from mast cells in TRX-tg mice is suppressed by inhibition of ROS generation. As ROS are involved in mast cell activation and facilitate mediator release, TRX may be a key signaling molecule regulating the early events in the IgE signaling in mast cells and the allergic inflammation.展开更多
Human placenta-derived mononuclear cells （MNC） were isolated by a Percoll density gradient and cultured in mesenchymal stem cell （MSC） maintenance medium. The homogenous layer of adherent cells exhibited a typical...Human placenta-derived mononuclear cells （MNC） were isolated by a Percoll density gradient and cultured in mesenchymal stem cell （MSC） maintenance medium. The homogenous layer of adherent cells exhibited a typical fibroblastlike morphology, a large expansive potential, and cell cycle characteristics including a subset of quiescent cells. In vitro differentiation assays showed the tripotential differentiation capacity of these cells toward adipogenic, osteogenic and chondrogenic lineages. Flow cytometry analyses and immunocytochemistry stain showed that placental MSC was a homogeneous cell population devoid of hematopoietic cells, which uniformly expressed CD29, CD44, CD73, CD105, CD166, laminin, fibronectin and vimentin while being negative for expression of CD31, CD34, CD45 and m-smooth muscle actin. Most importantly, immuno-phenotypic analyses demonstrated that these cells expressed class Ⅰ major histocompatibility complex （MHC-I）, but they did not express MHC-Ⅱ molecules. Additionally these cells could suppress umbilical cord blood （UCB） lymphocytes proliferation induced by cellular or nonspecific mitogenic stimuli. This strongly implies that they may have potential application in allograft transplantation. Since placenta and UCB are homogeneous, the MSC derived from human placenta can be transplanted combined with hematopoietic stem cells （HSC） from UCB to reduce the potential graft-versus-host disease （GVHD） in recipients.展开更多
Some of the most common human cancers, including breast cancer, prostate cancer, and lung cancer, metastasize with avidity to bone. What is the basis for their preferential growth within the bone microenvironment? Bid...Some of the most common human cancers, including breast cancer, prostate cancer, and lung cancer, metastasize with avidity to bone. What is the basis for their preferential growth within the bone microenvironment? Bidirectional interactions between tumor cells and cells that make up bone result in a selective advantage for tumor growth and can lead to bone destruction or new bone matrix deposition. This review discusses our current understanding of the molecu- lar components and mechanisms that are responsible for those interactions.展开更多
Spontaneous Ca2+ oscillations in vascular smooth muscle cells have been modeled using a single Ca2+ pool. This report describes spontaneous Ca2+ oscillations dependent on two separate Ca2+ sources for the nuclear vers...Spontaneous Ca2+ oscillations in vascular smooth muscle cells have been modeled using a single Ca2+ pool. This report describes spontaneous Ca2+ oscillations dependent on two separate Ca2+ sources for the nuclear versus cytoplas- mic compartments. Changes in free intracellular Ca2+ were monitored with ratiometric Ca2+- fluorophores using confo- cal microscopy. On average, spontaneous oscillations developed in 79% of rat aortic smooth muscle cells that were synchronous between the cytoplasm and nucleus. Reduction of extracellular Ca2+ (< 1 μM) decreased the frequency and amplitude of the cytoplasmic oscillations with 48% of the oscillations asynchronous between the nuclear and cytoplasmic compartments. Similar results were obtained with the Ca2+ channel blockers, nimodipine and diltiazem. Arg-vasopressin (AVP) induced a rapid release of intracellular Ca2+ stores that was greater in the nuclear compartment (4.20 ± 0.23 ratio units, n = 56) than cytoplasm (2.54 ± 0.28) in cells that had spontaneously developed prior oscillations. Conversely, cells in the same conditions lacking oscillations had a greater AVP-induced Ca2+ transient in the cytoplasm (4.99 ± 0.66, n = 17) than in the nucleus (2.67 ± 0.29). Pre-treatment with Ca2+ channel blockers depressed the AVP responses in both compartments with the cytoplasmic Ca2+ most diminished. Depletion of internal Ca2+ stores prior to AVP exposure blunted the nuclear response, mimicking the response of cells that lacked prior oscillations. Spontaneous oscillating cells had a greater sarcoplasmic reticulum network than cells that did not oscillate. We propose that sponta- neous nuclear oscillations rely on perinuclear sarcoplasmic reticulum stores, while the cytoplasmic oscillations rely on Ca2+ influx.展开更多
Kinetics of thymocyte development in vivo during embryogenesis was pursued. The early development of thymocytes in the fetal and neonatal BALB/c mice was discontinuous, with four waves of cell proliferation occurring ...Kinetics of thymocyte development in vivo during embryogenesis was pursued. The early development of thymocytes in the fetal and neonatal BALB/c mice was discontinuous, with four waves of cell proliferation occurring at fetal day (Fd) 14 to 17, Fd 18 to day (D) 1 after birth, D 2 to D 5 and D6 thereafter. The first three proliferation waves coincided with the generation of CD4/'CD8/' (DP), TCR+CD4/hiCD8-/lo (CD4 SP), and TCR+CD4-/loCD8int/hi (CD8 SP) thymocytes, respectively. The transition from DN to DP cells was further investigated and it was found out that there were two differential pathways via immature single positive (ISP) cells in the BALB/c mice, each functioning at different fetal ages. One is via TCR-CD4-CD8+ cells, occurring between Fd 15 and Fd 17 and the other is via TCR-CD4+CD8- cells, occurring from Fd 17 until birth. In contrast, the TCR-CD4-CD8+ pathway dominated overwhelmingly in the C57BL/6 mice. These findings shed new light on the hypothesis that the differential pathway preference varies with mouse strains. With respect to the shift in the intensity of CD4 and CD8 expression on thymocytes from fetal to adult mice, the TCR+CD4/hiCD8-/lo, and TCR+CD4-/loCD8int/hi subsets might be equivalent to the medullary type TCR+CD4/CD8 SP cells.展开更多
This is the First Symposium on Frontiers of Biomedical Science organized by Cell Research, a peerreviewed journal supported by the Chinese Academy of Sciences. In recent years, epigenetics have shown its growing impor...This is the First Symposium on Frontiers of Biomedical Science organized by Cell Research, a peerreviewed journal supported by the Chinese Academy of Sciences. In recent years, epigenetics have shown its growing importance for the understanding of life processes and the control of human diseases in the genome era. I do hope that this timely symposium展开更多
The Chinese Medical Journal(CMJ)is published monthly in English by the Chinese Medical Association,and is a peer reviewed general medical Journal for all doctors,researchers,and health workers regardlessof their medic...The Chinese Medical Journal(CMJ)is published monthly in English by the Chinese Medical Association,and is a peer reviewed general medical Journal for all doctors,researchers,and health workers regardlessof their medical specialty or type of employment.Established in 1887,it is the oldest medical展开更多
Apoptosis is a form of genetically programmed cell death, which plays a key role in regulation of cellularity in a variety of tissue and cell types including the cardiovascular tissues. Under both physiological and pa...Apoptosis is a form of genetically programmed cell death, which plays a key role in regulation of cellularity in a variety of tissue and cell types including the cardiovascular tissues. Under both physiological and pathophysiological conditions, various biophysiological and biochemical factors, including mechanical forces, reactive oxygen and nitrogen species, cytokines, growth factors, oxidized lipoproteins, etc., may influence apoptosis of vascular cells. The Fas/Fas ligand/caspase death-signaling pathway, Bcl-2 protein family/mitochondria, the tumor suppressive gene p53, and the proto-oncogene c-myc may be activated in atherosclerotic lesions, and mediates vascular apoptosis during the development of atherosclerosis. Abnormal expression and dysfunction of these apoptosis-regulating genes may attenuate or accelerate vascular cell apoptosis and affect the integrity and stability of atherosclerotic plaques. Clarification of the molecular mechanism that regulates apoptosis may help design a new strategy for treatment of atherosclerosis and its major complication, the acute vascular syndromes.展开更多
GABA transporter 1(GAT1) takes important roles in multiple physiological processes through the uptake and release of GABA, but the regulation of GAT1 gene expression in different tissues is rarely known. To address th...GABA transporter 1(GAT1) takes important roles in multiple physiological processes through the uptake and release of GABA, but the regulation of GAT1 gene expression in different tissues is rarely known. To address the question, first, 5’ Rapid amplification of cDNA end (RACE) was used to determine GAT1 transcriptional starting sites in neonatal mouse cerebral cortex and intestine, adult mouse brain and adult rat testis. The products of 5’RACE were confirmed by DNA sequencing. We found that the transcript of GAT1 in neonatal mouse cerebral cortex and adult mouse brain starts at the same site (inside of exon 1), while in mouse intestine, GAT1 starts transcription in intron 1, and in rat testis, the transcript of GAT1 has an additional untranslation exon to the 5’ direction.展开更多
Human μ-opioid receptor （HμOR） with a tag of six consecutive histidines at its carboxyl terminus had been expressed in recombinant baculovirus infected Sf9 insect cells.The maximal binding capacity for the [3H] di...Human μ-opioid receptor （HμOR） with a tag of six consecutive histidines at its carboxyl terminus had been expressed in recombinant baculovirus infected Sf9 insect cells.The maximal binding capacity for the [3H] diprenorphine and [3H]ohmefentanyl （Ohm） were 9.1± 0.7 and 6.52±0.23 nmol/g protein, respectively. The [3H] diprenorphine or [3H] Ohm binding to the receptor expressed in Sf9 cells was strongly inhibited by μ-selective agonists [D-Ala2], N-methylPhe4, glyol5]enkephalin （DAGO）, Ohm, and morphine, but neither by δ nor by K selective agonist. Na+ （100 mM） and GTP （50 μM） could reduce HμOR agonists etorphine and Ohm affinity binding to the overexpressed HμOR. μ-selective agonists DAGO and Ohm effectively stimulated [35S]GTPγS binding (EC50 = 2.7nM and 6.9 nM) and inhibited forskolin- stimulated cAMP accumulation (IC50 = 0.9 nM and 0.3 nM). The agonist-dependent effects could be blocked by opioid antagonist naloxone or by pretreatment of cells with pertussis toxin （PTX）. These results demonstrated that HμOR overexpressed in Sf9 insect cells functionally coupled to endogenous Gi/o proteins.展开更多
FT Ⅰ (AAAAGGGGAAGCAGAG), a poly purine ele-ment within the myloid-lineage specific enhancer (En 1) of the mouse myeloperoxidase gene [1, 2] has been fur-ther characterised. 1, FT Ⅰ functions as a myeloid-lineage spe...FT Ⅰ (AAAAGGGGAAGCAGAG), a poly purine ele-ment within the myloid-lineage specific enhancer (En 1) of the mouse myeloperoxidase gene [1, 2] has been fur-ther characterised. 1, FT Ⅰ functions as a myeloid-lineage specific transcription regulatory element; 2, WEHI 3BD+ cells have higher binding activity to FT Ⅰ and express the proteins which could form the unique DNA-protein com-plex(es) of FT Ⅰ;. 3, The essential sequence for the specific DNA-protein interactions of FT Ⅰ is AAAAGGGGAAGC; 4, South-western analysis in conjunction with the compe-tition assay of the proteins binding to FT Ⅰ, has revealed a 28 kd protein in WEHI 3BD+ cells that displays the properties of the putative transcription factor which acts through FT Ⅰ. These new findings have demonstrated both the functional myeloid-lineage specificity and the novelty of FT Ⅰ.展开更多
The distribution of acetylcholinesterase(AChE)-positive structures in the developing rat spinal cord was studied with AChE-histochemistry.AChE-positive perikarya were first seen on embryonic day 14(E14) in the ventrol...The distribution of acetylcholinesterase(AChE)-positive structures in the developing rat spinal cord was studied with AChE-histochemistry.AChE-positive perikarya were first seen on embryonic day 14(E14) in the ventrolateral portion of the spinal cord.From that time onward.AChE=containing cells appeared gradually in the intermediate gray,dorsal horn and lateral spinal nucleus of the spinal cord in a ventral-to-dorsal,and lateral-to-medial order.No obvious rostral-to-caudal sequence was found.At birth,the distribution pattern of AChE-positive perikarya was basically similar to that in adults.After birth a dramatic increase in the AChE staining intensity extended from postnatal day 5(P5) to postnatal day 21(P21),In addition,two phases of transient AChE staining were observed in the external surface of the dorsal horn from embryonic day 15(E15) to embryonic day 21(E21) and in the marginal layer from embryonic day 21(E21) to postnatal day 14(P14),respectively.展开更多
Found in all metazoans, microRNAs or miRNAs are small non-coding RNA of-22 nucleotides in length that completely reshaped our understanding of gene regulation. This new class ofgene regulator is mostly transcribed by ...Found in all metazoans, microRNAs or miRNAs are small non-coding RNA of-22 nucleotides in length that completely reshaped our understanding of gene regulation. This new class ofgene regulator is mostly transcribed by the RNA polymerase II producing a long stem-loop structure, called primary- or pri-miRNA, that will first be processed in the cell nucleus by a multiprotein complex called microprocessor to generate a shorter RNA structure called precursor- or pre-miRNA.展开更多
Abnormal activity of an endogenous retroviral promoter plays a critical role in the development of Hodgkin＇s lymphoma, according to a paper in the May issue of Nature Medicine [ 1 ]. Throughout evolution, infection o...Abnormal activity of an endogenous retroviral promoter plays a critical role in the development of Hodgkin＇s lymphoma, according to a paper in the May issue of Nature Medicine [ 1 ]. Throughout evolution, infection ofgermline cells with retroviruses has led to the progressive accumulation of endogenous retroviral elements in genomes. The infecting retroviral RNA is reverse transcribed into DNA which inserts into the genome and is then transmitted to offspring in a Mendelian manner.展开更多
文摘Polycomb group genes play crucial roles in the maintenance of the transcriptionally silenced state of genes for proper cell differentiation in animals and plants. While components of the polycomb repressive complex2 （PRC2） are evolutionarily conserved and their functions are extensively studied in plants, PRCI differs considerably between animals and plants, and its functions in plants are as yet not well described. Previous studies have identified the Arabidopsis AtRINGla and AtRINGlb as homologues of the animal PRC1 subunit RING1. Here, we show that the Atringla Atringlb double mutant exhibits derepression of embryonic traits during vegetative growth. Accordingly, several key regulatory genes involved in embryogenesis and stem cell activity are ectopically expressed in the mutant. Furthermore, we show that the mutant phenotypes and increased expression of regulatory genes are enhanced by the PRC2 mutant c/f. Finally, we show that three homologues of the animal PRCl-subunit ring-finger protein BMI1, AtBMIIa, AtBMIlb and AtBMIlc, can bind with AtRINGla or AtRINGIb, and in addition, AtBMIlc can bind with LHP1. The Atbmila Atbmilb double mutant shows derepression of embryonic traits similar to that of the Atringla Atringlb double mutant. Interestingly, expression levels of AtBMIla, AtBMIlb and AtBMIlc are elevated in the Atringla Atringlb mutant and those of AtBMIlc, AtRINGla and AtRINGlb are elevated in the Atbmila Atbmilb mu- tant, suggesting a self-regulatory feedback mechanism. Taken together, our results illuminate crucial functions of the PRCl-like ring-finger components in stable repression of embryonic traits and regulatory genes for proper somatic growth.
文摘Endocytosis occurs at the cell surface and involves internalization of the plasma membrane （PM） along with its constituent membrane proteins and lipids. Endocytosis is involved in sampling of the extracellular milieu and also serves to regulate various processes initiated at the cell surface. These include nutrient uptake, signaling from cell- surface receptors, and many other processes essential for cell and tissue functioning in metazoans. It is also central to the maintenance of PM lipid and protein homeostasis. There are multiple means of internalization that operate concurrently, at the cell surface. With advancement in high-resolution visualization techniques, it is now possible to track multiple endocytic cargo at the same time, revealing a remarkable diversity of endocytic processes in a single cell. A combination of live cell imaging and efficient genetic manipulations has also aided in understanding the functional hierarchy of molecular players in these mechanisms of internalization. Here we provide an account of various endocytic routes, their mechanisms of operation and occurrence across phyla.
文摘Drosophila RYBP （dRYBP; Ringl and Y_Y1 Binding Protein） is a Polycomb and trithorax interacting protein, whose homologous RYBP/DEDAF mammalian counterparts exhibit tumor cell-specific killing activity. Here we show that although endogenous dRYBP is not involved in developmental apoptosis, high levels of exogenous dRYBP induce apoptosis in all the imaginal discs of the fly, indicating that dRYBP apoptotic activity is not specific to tumor cells. We also show that dRYBP-induced apoptosis is inhibited by high levels of either p35 or DIAP1 （Drosophila Inhibitor of Apoptosis Protein 1）, and requires the function of the pro-apoptotic REAPER, HID and GRIM proteins, the apical caspase DREDD, the adaptor dFADD protein as well as TRITHORAX （TRX）, an epigenetic transcriptional regulator. Furthermore, we demonstrate that overexpression of TRX also induces apoptosis in the imaginal discs. Finally, we show that the expression of reaper-lacZ is upregulated both upon dRYBP-induced apoptosis and upon TRX- induced apoptosis in imaginal discs and that the reaper gene is a direct target of dRYBP in Drosophila embryos. Our results indicate that dRYBP triggers in a receptor-mediated apoptotic pathway that also includes TRX-dependent epigenetic regulation of gene expression.
文摘While severe acute respiratory syndrome coronavirus （SARS-CoV）~as initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that yirus entry may also involve endocytosis. We have found that SARS-CoV enters cells viapH- and receptor-dependent endocytosis. Treatment of cells with either SARS-COV spike protein or spike-bearing pseudoviruses resulted in the translocation of angiotensin-converting enzyme 2 （ACE2）, the functional receptor of SARS-CoV, from the cell surface to endosomes. In addition, the spike-bearing pseudoviruses and early endosome antigen 1 were found to colocalize in endosomes. Further analyses using specific endocytic path- way inhibitors and dominant-negative Epsl5 as well as caveolin-1 colocalization study suggested that virus entry was mediated by a clathrin- and caveolae-independent mechanism. Moreover, cholesterol- and sphingolipid-rich lipid raft microdomains in the plasma membrane, which have been shown to act as platforms for many physiological signaling pathways, were shown to be involved in virus entry. Endocytic entry of SARS-CoV may expand the cellular range of SARS-CoV infection, and our findings here contribute to the understanding of SARS-CoV pathogenesis, providing new information for anti-viral drug research.
文摘The transcription factor p53 is a key regulator of the DNA damage response to genotoxic stress in higher eukaryotes. Mutation/inactivation of p53 has been implicated in the pathogenesis of many tumor types [ 1 ]. p53 responds to cellular insults, such as DNA damage or hypoxia, either by arresting the cell cycle so that DNA damage could be repaired, or by triggering apoptosis ifDNA repair is futile. Transcriptional activation of apoptosis-related genes by p53 is critical for the induction of programmed cell death. In search of target genes of p53 involved in cell-cycle arrest and/or apoptosis, numerous p53-inducible genes have been identified over the past 15 years. Among them are genes such as Bax [2], Puma [3], Noxa [4], Bid [5], p21waf1 [6], and many others. These apoptosis-related factors function in different cell organelles including the cytosolic membrane, the mitochondria and the cytosol. However, none of these candidate effectors alone can be accountable for the complicated mechanisms underlying the p53 transcription-dependent apoptotic signaling pathways. Earlier work implicated that Bad may contribute to p53-induced mitochondria-mediated apoptosis, yet no known molecular mechanism had been ascribed to this observation. Our recent study published in Molecular and Cellular Biology revealed that a functional p53-binding element exists at the human bad genomic locus, residing roughly 6.6 kb upstream of the Bad translation start codon [7]. The fact that this p53-binding element is relatively far away from the Bad promoter could be the reason why it has not been recognized as a p53-binding element until today. Interestingly enough, we found that a similar p53-binding region is well conserved approximately 7 kb upstream of the murine bad locus （our unpublished data）.
文摘The application of genomic technologies to the study of mRNA processing is increasingly conducted in metazoan organisms in order to understand the complex events that occur during and after transcription. Large-scale systems analyses ofmRNA-protein interactions and mRNA dynamics have revealed specificity in mRNA transcription, splicing, transport, translation, and turnover, and have begun to make connections between the different layers ofmRNA processing. Here, we review global studies of post-transcriptional processes and discuss the challenges facing our understanding of mRNA regulation in metazoan organisms. In parallel, we examine genome-scale investigations that have expanded our knowledge of RNA-binding proteins and the networks of mRNAs that they regulate.
文摘Thioredoxin-1 （TRX） is a stress-inducible redox-regulatory protein with antioxidative and anti-inflammatory effects. Here we show that the release of histamine from mast cells elicited by cross-linking of high-affinity receptor for IgE （FcεRI） was significantly suppressed in TRX transgenic （TRX-tg） mice compared to wild type （WT） mice. Intracellular reactive oxygen species （ROS） of mast cells stimulated by IgE and antigen was also reduced in TRX-tg mice compared to WT mice. Whereas there was no difference in the production ofcytokines （IL-6 and TNF-α） from mast cells in response to 2,4-dinitrophenylated bovine serum albumin （DNP-BSA） stimulation in TRX-tg and WT mice. Immunological status of TRX-tg mice inclined to T helper （Th） 2 dominant in primary immune response, although there was no difference in the population of dendritic cells （DCs） and regulatory T cells. We conclude that the histamine release from mast cells in TRX-tg mice is suppressed by inhibition of ROS generation. As ROS are involved in mast cell activation and facilitate mediator release, TRX may be a key signaling molecule regulating the early events in the IgE signaling in mast cells and the allergic inflammation.
基金This study was supported by a grant from National Natural Science Foundation of China(No.30271245)Hi-Tech Research and Development Program of China(863 Program)(No.2003AA205170)+1 种基金National Basic Research Program of China(973 Program)(No.G 1999054302)a grant from Bejing Gynecology and Obstetrics Hospital Affiliate of Capital University of Medical Sciences.
文摘Human placenta-derived mononuclear cells （MNC） were isolated by a Percoll density gradient and cultured in mesenchymal stem cell （MSC） maintenance medium. The homogenous layer of adherent cells exhibited a typical fibroblastlike morphology, a large expansive potential, and cell cycle characteristics including a subset of quiescent cells. In vitro differentiation assays showed the tripotential differentiation capacity of these cells toward adipogenic, osteogenic and chondrogenic lineages. Flow cytometry analyses and immunocytochemistry stain showed that placental MSC was a homogeneous cell population devoid of hematopoietic cells, which uniformly expressed CD29, CD44, CD73, CD105, CD166, laminin, fibronectin and vimentin while being negative for expression of CD31, CD34, CD45 and m-smooth muscle actin. Most importantly, immuno-phenotypic analyses demonstrated that these cells expressed class Ⅰ major histocompatibility complex （MHC-I）, but they did not express MHC-Ⅱ molecules. Additionally these cells could suppress umbilical cord blood （UCB） lymphocytes proliferation induced by cellular or nonspecific mitogenic stimuli. This strongly implies that they may have potential application in allograft transplantation. Since placenta and UCB are homogeneous, the MSC derived from human placenta can be transplanted combined with hematopoietic stem cells （HSC） from UCB to reduce the potential graft-versus-host disease （GVHD） in recipients.
文摘Some of the most common human cancers, including breast cancer, prostate cancer, and lung cancer, metastasize with avidity to bone. What is the basis for their preferential growth within the bone microenvironment? Bidirectional interactions between tumor cells and cells that make up bone result in a selective advantage for tumor growth and can lead to bone destruction or new bone matrix deposition. This review discusses our current understanding of the molecu- lar components and mechanisms that are responsible for those interactions.
文摘Spontaneous Ca2+ oscillations in vascular smooth muscle cells have been modeled using a single Ca2+ pool. This report describes spontaneous Ca2+ oscillations dependent on two separate Ca2+ sources for the nuclear versus cytoplas- mic compartments. Changes in free intracellular Ca2+ were monitored with ratiometric Ca2+- fluorophores using confo- cal microscopy. On average, spontaneous oscillations developed in 79% of rat aortic smooth muscle cells that were synchronous between the cytoplasm and nucleus. Reduction of extracellular Ca2+ (< 1 μM) decreased the frequency and amplitude of the cytoplasmic oscillations with 48% of the oscillations asynchronous between the nuclear and cytoplasmic compartments. Similar results were obtained with the Ca2+ channel blockers, nimodipine and diltiazem. Arg-vasopressin (AVP) induced a rapid release of intracellular Ca2+ stores that was greater in the nuclear compartment (4.20 ± 0.23 ratio units, n = 56) than cytoplasm (2.54 ± 0.28) in cells that had spontaneously developed prior oscillations. Conversely, cells in the same conditions lacking oscillations had a greater AVP-induced Ca2+ transient in the cytoplasm (4.99 ± 0.66, n = 17) than in the nucleus (2.67 ± 0.29). Pre-treatment with Ca2+ channel blockers depressed the AVP responses in both compartments with the cytoplasmic Ca2+ most diminished. Depletion of internal Ca2+ stores prior to AVP exposure blunted the nuclear response, mimicking the response of cells that lacked prior oscillations. Spontaneous oscillating cells had a greater sarcoplasmic reticulum network than cells that did not oscillate. We propose that sponta- neous nuclear oscillations rely on perinuclear sarcoplasmic reticulum stores, while the cytoplasmic oscillations rely on Ca2+ influx.
基金supported by grants from National 973 Program in China(No.G1999053904)National Natural Sciences Foundation of China(No.39730410)
文摘Kinetics of thymocyte development in vivo during embryogenesis was pursued. The early development of thymocytes in the fetal and neonatal BALB/c mice was discontinuous, with four waves of cell proliferation occurring at fetal day (Fd) 14 to 17, Fd 18 to day (D) 1 after birth, D 2 to D 5 and D6 thereafter. The first three proliferation waves coincided with the generation of CD4/'CD8/' (DP), TCR+CD4/hiCD8-/lo (CD4 SP), and TCR+CD4-/loCD8int/hi (CD8 SP) thymocytes, respectively. The transition from DN to DP cells was further investigated and it was found out that there were two differential pathways via immature single positive (ISP) cells in the BALB/c mice, each functioning at different fetal ages. One is via TCR-CD4-CD8+ cells, occurring between Fd 15 and Fd 17 and the other is via TCR-CD4+CD8- cells, occurring from Fd 17 until birth. In contrast, the TCR-CD4-CD8+ pathway dominated overwhelmingly in the C57BL/6 mice. These findings shed new light on the hypothesis that the differential pathway preference varies with mouse strains. With respect to the shift in the intensity of CD4 and CD8 expression on thymocytes from fetal to adult mice, the TCR+CD4/hiCD8-/lo, and TCR+CD4-/loCD8int/hi subsets might be equivalent to the medullary type TCR+CD4/CD8 SP cells.
文摘This is the First Symposium on Frontiers of Biomedical Science organized by Cell Research, a peerreviewed journal supported by the Chinese Academy of Sciences. In recent years, epigenetics have shown its growing importance for the understanding of life processes and the control of human diseases in the genome era. I do hope that this timely symposium
文摘The Chinese Medical Journal(CMJ)is published monthly in English by the Chinese Medical Association,and is a peer reviewed general medical Journal for all doctors,researchers,and health workers regardlessof their medical specialty or type of employment.Established in 1887,it is the oldest medical
文摘Apoptosis is a form of genetically programmed cell death, which plays a key role in regulation of cellularity in a variety of tissue and cell types including the cardiovascular tissues. Under both physiological and pathophysiological conditions, various biophysiological and biochemical factors, including mechanical forces, reactive oxygen and nitrogen species, cytokines, growth factors, oxidized lipoproteins, etc., may influence apoptosis of vascular cells. The Fas/Fas ligand/caspase death-signaling pathway, Bcl-2 protein family/mitochondria, the tumor suppressive gene p53, and the proto-oncogene c-myc may be activated in atherosclerotic lesions, and mediates vascular apoptosis during the development of atherosclerosis. Abnormal expression and dysfunction of these apoptosis-regulating genes may attenuate or accelerate vascular cell apoptosis and affect the integrity and stability of atherosclerotic plaques. Clarification of the molecular mechanism that regulates apoptosis may help design a new strategy for treatment of atherosclerosis and its major complication, the acute vascular syndromes.
基金foundations from Chinese Academy of Sciences and Special Funds for Major State Basic reseaxch of China (G1999053903).
文摘GABA transporter 1(GAT1) takes important roles in multiple physiological processes through the uptake and release of GABA, but the regulation of GAT1 gene expression in different tissues is rarely known. To address the question, first, 5’ Rapid amplification of cDNA end (RACE) was used to determine GAT1 transcriptional starting sites in neonatal mouse cerebral cortex and intestine, adult mouse brain and adult rat testis. The products of 5’RACE were confirmed by DNA sequencing. We found that the transcript of GAT1 in neonatal mouse cerebral cortex and adult mouse brain starts at the same site (inside of exon 1), while in mouse intestine, GAT1 starts transcription in intron 1, and in rat testis, the transcript of GAT1 has an additional untranslation exon to the 5’ direction.
文摘Human μ-opioid receptor （HμOR） with a tag of six consecutive histidines at its carboxyl terminus had been expressed in recombinant baculovirus infected Sf9 insect cells.The maximal binding capacity for the [3H] diprenorphine and [3H]ohmefentanyl （Ohm） were 9.1± 0.7 and 6.52±0.23 nmol/g protein, respectively. The [3H] diprenorphine or [3H] Ohm binding to the receptor expressed in Sf9 cells was strongly inhibited by μ-selective agonists [D-Ala2], N-methylPhe4, glyol5]enkephalin （DAGO）, Ohm, and morphine, but neither by δ nor by K selective agonist. Na+ （100 mM） and GTP （50 μM） could reduce HμOR agonists etorphine and Ohm affinity binding to the overexpressed HμOR. μ-selective agonists DAGO and Ohm effectively stimulated [35S]GTPγS binding (EC50 = 2.7nM and 6.9 nM) and inhibited forskolin- stimulated cAMP accumulation (IC50 = 0.9 nM and 0.3 nM). The agonist-dependent effects could be blocked by opioid antagonist naloxone or by pretreatment of cells with pertussis toxin （PTX）. These results demonstrated that HμOR overexpressed in Sf9 insect cells functionally coupled to endogenous Gi/o proteins.
文摘FT Ⅰ (AAAAGGGGAAGCAGAG), a poly purine ele-ment within the myloid-lineage specific enhancer (En 1) of the mouse myeloperoxidase gene [1, 2] has been fur-ther characterised. 1, FT Ⅰ functions as a myeloid-lineage specific transcription regulatory element; 2, WEHI 3BD+ cells have higher binding activity to FT Ⅰ and express the proteins which could form the unique DNA-protein com-plex(es) of FT Ⅰ;. 3, The essential sequence for the specific DNA-protein interactions of FT Ⅰ is AAAAGGGGAAGC; 4, South-western analysis in conjunction with the compe-tition assay of the proteins binding to FT Ⅰ, has revealed a 28 kd protein in WEHI 3BD+ cells that displays the properties of the putative transcription factor which acts through FT Ⅰ. These new findings have demonstrated both the functional myeloid-lineage specificity and the novelty of FT Ⅰ.
文摘The distribution of acetylcholinesterase(AChE)-positive structures in the developing rat spinal cord was studied with AChE-histochemistry.AChE-positive perikarya were first seen on embryonic day 14(E14) in the ventrolateral portion of the spinal cord.From that time onward.AChE=containing cells appeared gradually in the intermediate gray,dorsal horn and lateral spinal nucleus of the spinal cord in a ventral-to-dorsal,and lateral-to-medial order.No obvious rostral-to-caudal sequence was found.At birth,the distribution pattern of AChE-positive perikarya was basically similar to that in adults.After birth a dramatic increase in the AChE staining intensity extended from postnatal day 5(P5) to postnatal day 21(P21),In addition,two phases of transient AChE staining were observed in the external surface of the dorsal horn from embryonic day 15(E15) to embryonic day 21(E21) and in the marginal layer from embryonic day 21(E21) to postnatal day 14(P14),respectively.
文摘Found in all metazoans, microRNAs or miRNAs are small non-coding RNA of-22 nucleotides in length that completely reshaped our understanding of gene regulation. This new class ofgene regulator is mostly transcribed by the RNA polymerase II producing a long stem-loop structure, called primary- or pri-miRNA, that will first be processed in the cell nucleus by a multiprotein complex called microprocessor to generate a shorter RNA structure called precursor- or pre-miRNA.
文摘Abnormal activity of an endogenous retroviral promoter plays a critical role in the development of Hodgkin＇s lymphoma, according to a paper in the May issue of Nature Medicine [ 1 ]. Throughout evolution, infection ofgermline cells with retroviruses has led to the progressive accumulation of endogenous retroviral elements in genomes. The infecting retroviral RNA is reverse transcribed into DNA which inserts into the genome and is then transmitted to offspring in a Mendelian manner.
