Mitochondrial damage is closely related to the occurrence of many diseases.However,accurate monitoring and reporting of mitochondrial damage are not easy.Here,we developed a small molecule fluorescent probe named CB-C...Mitochondrial damage is closely related to the occurrence of many diseases.However,accurate monitoring and reporting of mitochondrial damage are not easy.Here,we developed a small molecule fluorescent probe named CB-Cl,which has splendid spectral properties(large Stokes shift,strong affinity for RNA,etc.)and excellent targeting ability to intracellular mitochondria.After mitochondria were damaged by external stimuli,CB-Cl would light up the nucleolus as a signal reporter.The cascade imaging of mitochondria and nucleolus using CB-Cl can monitor and visualize the mitochondrial status in living cells in real-time.Based on the above advantages,the probe CB-Cl has reference significance for the related research of mitochondrial damage and the prevention and treatment of related diseases.展开更多
By using the DNA specific cytochemical staining method (NAMA_Ur) and conventional electron microscopic technique, the authors examined the configuration of intranucleolar DNA in Allium cepa L. cells and found that...By using the DNA specific cytochemical staining method (NAMA_Ur) and conventional electron microscopic technique, the authors examined the configuration of intranucleolar DNA in Allium cepa L. cells and found that nucleolar DNA within the fibrillar center (FC) underwent a structural transformation process from condensed to extended state. The authors' observations also displayed a continuous arrangement process of nucleolar DNA, i.e., the extranucleolar DNA entered FC through the nucleolar organizer region (NOR) channel, then extended to the periphery of FC or to the border between FC and dense fibrillar component (DFC), and distributed along the periphery of FC. Thence, by passing through the NOR channel between FCs, the nucleolar DNA continued to transfer to other FCs and arranged in the same above_mentioned forms.展开更多
Processing of pre-rRNA is one of the major events taking place In the nucleolus. U3 snoRNA, an rRNA spliceosomal factor, is suggested to be essential in the first cleavage step of the 5' ETS sequence in the proces...Processing of pre-rRNA is one of the major events taking place In the nucleolus. U3 snoRNA, an rRNA spliceosomal factor, is suggested to be essential in the first cleavage step of the 5' ETS sequence in the processing of pre-rRNA. Identification of U3 in the nucleolus provides a piece of indirect evidence for pre-rRNA processing site and transportation of processing products. In the present study, subnucleolar distribution of U3 snoRNA in the nucleolus of Pisum sativum L. was studied by in situ hybridization with a U3 snoRNA probe. The results showed that the U3 labeling signals were distributed throughout dense fibrillar components (DFCs) and granular components (GCs), while no signal was found in fibrillar centers (FCs). When treated with actinomycine D (AMD), the labeling signals were decreased. Along with the increase of the AMD treatment time, the labeling signals became fewer and they were found in the distal regions of DFC and GC. Our results indicated that pre-rRNA splicing took place in the regions of DFC and GC, and the transportation of pre-rRNA processing products was from the regions around FCs towards the distal regions.展开更多
The nucleolus is the most prominent compartment in the nucleus and known as the site for ribosome biogenesis in eucaryotes. In contrast, there is no such equivalent structure for ribosome synthesis in procaryotes. Thi...The nucleolus is the most prominent compartment in the nucleus and known as the site for ribosome biogenesis in eucaryotes. In contrast, there is no such equivalent structure for ribosome synthesis in procaryotes. This raises two concerns that how does the nucleolus evolve and that whether the nucleolus remains playing a single role in ribosome biogenesis along the evolution. Increasing data support new nucleolus functions, including signal recognition particle assembly, small RNA modification, telomerase maturation, cell-cycle and aging control, and cell stress sensor. Multiple functions of the nucleolus possibly result from the plurifunctionality of nucleolar proteins, such as nucleolin and Nopp 140. Proteomic analyses of human and Arabidopsis nucleolus lead a remarkable progress in understanding the evolution and new functions of nucleoli. In this review, we present a brief history of nucleolus research and new concepts and unresolved questions. Also, we introduce hepatitis D virus for studying the communication between the nucleolus and other subnuclear compartments, and Caenorhabditis elegans for the role of nucleolus in the development and the epistatic control of nucleologenesis.展开更多
The nucleolus,as the‘nucleus of the nucleus’,is a prominent subcellular organelle in a eukaryocyte.The nucleolus serves as the centre for ribosome biogenesis,as well as an important site for cell-cycle regulation,ce...The nucleolus,as the‘nucleus of the nucleus’,is a prominent subcellular organelle in a eukaryocyte.The nucleolus serves as the centre for ribosome biogenesis,as well as an important site for cell-cycle regulation,cellular senescence,and stress response.The protein composition of the nucleolus changes dynamically through protein turnover to meet the needs of cellular activities or stress responses.Recent studies have identified a nucleolus-localized protein degradation pathway in zebrafish and humans,namely the Def-CAPN3 pathway,which is essential to ribosome production and cell-cycle progression,by controlling the turnover of multiple substrates(e.g.,ribosomal small-subunit[SSU]processome component Mpp10,transcription factor p53,check-point proteins Chk1 and Wee1).This pathway relies on the Ca2þ-dependent cysteine proteinase CAPN3 and is independent of the ubiquitin-mediated proteasome pathway.CAPN3 is recruited by nucleolar protein Def from cytoplasm to nucleolus,where it proteolyzes its substrates which harbor a CAPN3 recognition-motif.Def depletion leads to the exclusion of CAPN3 and accumulation of p53,Wee1,Chk1,and Mpp10 in the nucleolus that result in cell-cycle arrest and rRNA processing abnormality.Here,we summarize the discovery of the Def-CAPN3 pathway and propose its biological role in cell-cycle control and ribosome biogenesis.展开更多
The nucleolus is a subnuclear structure of eukaryocytes. It was thought that nucleolus only participates in the biogenesis and processing of rRNA. However, more and more evidence shows that it has many other functions...The nucleolus is a subnuclear structure of eukaryocytes. It was thought that nucleolus only participates in the biogenesis and processing of rRNA. However, more and more evidence shows that it has many other functions, such as tRNA precursor processing, stress sensing and it is also involved in gene silencing, senescence and cell cycle regulation. Here, we summarize the recent understandings about the nucleolar functions, the regulation of nucleolar localization of proteins and the role that the nucleolus plays in virus infection, in which some related studies of Herpes simplex virus type 1 (HSV-1) US11, UL24 and bovine herpesvirus-1 infected cell protein 27 (BICP27) carried out in our lab will also be included.展开更多
The effects of different concentrations of cadmium chloride ranging from 0.5 to 20 ppm on the nucleolus in root tip cells of Allium cepa were studied using the silver staining technique. The results indicated that aft...The effects of different concentrations of cadmium chloride ranging from 0.5 to 20 ppm on the nucleolus in root tip cells of Allium cepa were studied using the silver staining technique. The results indicated that after the treatment with Cd￣(2+),different changes in nucleolar morphology appeared. The nucleolar material was extruded from the nucleus into the cytoplasm,and the nucleoli at mitotic metaphase did not disappear. Apparently,cadmium showed a specific effects on the nucleoli in root tip cells of Allium cepa. The possible mechanism behind this phenomenon is also briefly discussed.展开更多
In our conversation with Prof.CHEN Lingling on the"dark mat ter"in the genome,namely the non-coding RNAs,we got to know two classes of long non-coding RNAs(lncRNAs)in close association with Prader-Willi synd...In our conversation with Prof.CHEN Lingling on the"dark mat ter"in the genome,namely the non-coding RNAs,we got to know two classes of long non-coding RNAs(lncRNAs)in close association with Prader-Willi syndrome(PWS).Each with a snoRNA cap on its either end,they fall in the sno-lncRNA family,a group of lncRNAs identified by CHEN’s group in their earlier research.Here we are meeting another member of the family,SLERT,a rescuer展开更多
We observed the ultrastructure of nucleolus in rat liver cells by conventional electron microscopy, and employed cytochemistry NAMA-Ur DNA specific stain method to analyze the distribution and position of nucleolar DN...We observed the ultrastructure of nucleolus in rat liver cells by conventional electron microscopy, and employed cytochemistry NAMA-Ur DNA specific stain method to analyze the distribution and position of nucleolar DNA in situ. The results showed that nucleolar DNA of rat liver cells comes from nucleolus-associated chromatin, and continuously extends in the dense fibrillar component (DFC) of nucleolus, localizes at the periphery of fibrillar center (FC) and in DFC. Furthermore, by employing anti-DNA/RNA hybrid antibodies, we directly and selectively labeled transcription sites of rRNA genes and testified that localization of transcription sites not only to DFC but also to the periphery of FC.展开更多
The location and configuration of nucleolar DNA have not been determined for a long time. In this paper, we have observed the nucleolar infrastructure and the character of nucleolar DNA in Allium cepa cells by convent...The location and configuration of nucleolar DNA have not been determined for a long time. In this paper, we have observed the nucleolar infrastructure and the character of nucleolar DNA in Allium cepa cells by conventional electron microscopy and the cytochemical NAMA-Ur DNA specific staining method. Furthermore, we have properly improved the NAMA-Ur method so as to analyze the location and configuration of nucleolar DNA in situ. Our results indicated that the nucleolar DNA in Allium cepa cells is mainly located at the border between fibrillar centers and dense fibribllar component, especially distributed in the configuration of encircling the fibrillar centers.展开更多
Microalgae are used in food and feed,and they are considered a potential feedstock for sustainably pro-duced chemicals and biofuel.However,production of microalgal-derived chemicals is not yet economically feasible.Ge...Microalgae are used in food and feed,and they are considered a potential feedstock for sustainably pro-duced chemicals and biofuel.However,production of microalgal-derived chemicals is not yet economically feasible.Genetic engineering could bridge the gap to industrial application and facilitate the production of novel products from microalgae.Here,we report the discovery of a novel gene expression system in the oleaginous microalga Nannochloropsis that exploits the highly efficient transcriptional activity of RNA po-lymerase I and an internal ribosome entry site for translation.We identified the nucleolus as a genomic safe harbor for Pol I transcription and used it to construct transformant strains with consistently strong trans-gene expression.The new expression system provides an outstanding tool for genetic and metabolic en-gineering of microalgae and thus will probably make substantial contributions to microalgal research.展开更多
Nucleolus organizer regions (Nor loci) of a range of Chinese wheat landraces and cultivars (Triticum aestivum L. em Thell.) were analysed using genomic DNA extracted from leaves. Only two allelic variants of the Nor-B...Nucleolus organizer regions (Nor loci) of a range of Chinese wheat landraces and cultivars (Triticum aestivum L. em Thell.) were analysed using genomic DNA extracted from leaves. Only two allelic variants of the Nor-B1 locus were found on chromosome 1B (Nor-B1a and Nor-B1g), while Nor-B1g was probably introduced from North America in the early 1960s. The even more recent introduction of the rye allele Nor-R1 in the early 1980s was also revealed. Eight allelic variants of the Nor-B2 locus on chromosome 6B (Nor-B2a, b, d, f, h, o, p and s) were identified. A Chinese origin for the a, d, f, o, p and s alkies is evident although the d allele was successfully introduced into Australian wheats in the early 1900s. Nor-B2h and Nor-B2b are again very recent introductions into Chinese wheat breeding programs, the former from CIMMYT wheats and the latter in association with the introduction of the 1RS/1BL translocation from Europe. On the basis of the presence of different combinations of Nor-B1 and Nor-B2 alleles, the Chinese wheat cultivars can be divided into eight phenotypic groups.展开更多
Nucleolus is the most prominent subnuclear structure, which performs a wide variety of functions in the eukaryotic cellular processes. In order to understand the structural and functional role of the nucleoli in bovin...Nucleolus is the most prominent subnuclear structure, which performs a wide variety of functions in the eukaryotic cellular processes. In order to understand the structural and functional role of the nucleoli in bovine cells, we analyzed the proteomic composition of the bovine nucleoli. The nucleoli were isolated from Madin Darby bovine kidney cells and subjected to proteomic analysis by LC-MS/MS after fractionation by SDS-PAGE and strong cation exchange chromatography. Analysis of the data using the Mascot database search and the GPM database search identified 311 proteins in the bovine nucleoli, which contained 22 proteins previously not identified in the proteomic analysis of human nucleoli. Analysis of the identified proteins using the GoMiner software suggested that the bovine nucleoli contained proteins involved in ribosomal biogenesis, cell cycle control, transcriptional, translational and post-translational regulation, transport, and structural organization.展开更多
Our conventional EM observations indicate that the nucleolus of Allium cepa is composedof the fibrillar centre (FC), fibrillar component (F) and granular region (G). FC is an elec-tron-lucid zone which contains conden...Our conventional EM observations indicate that the nucleolus of Allium cepa is composedof the fibrillar centre (FC), fibrillar component (F) and granular region (G). FC is an elec-tron-lucid zone which contains condensed or loosened chromatin. F is a circular, highly elec-tron-dense component which surrounds FC and consists of compact fibrils. EM autoradiographic(EM ARG) studies with ~3H- UdR reveal that the synthesis of ribosomal RNA (rRNA) takesplace in F. G is situated around F and composed of granules about 25 to 30 nm in diameter.EM ARG studies with ~3H- TdR demonstrate that silver grains are predominantly located in Gof the nucleolar periphery and the region of the nucleolus-associated chromatin. In addition,~3H- TdR labels are also present, but with a much lower frequency, in FC. F and G of the nu-cleolar interior. According to the distribution and sphere of silver grains in different compo-nents of the nucleolus, we suggest that the replication sites of ribosomal DNA (rDNA) aremainly located in G of the nucleolar periphery and the region of the nucleolus-associated chro-matin. Our observations also suggest that some segments of rDNA initiate their replication inFC and then move to the nucleolar periphery.展开更多
The CRISPR/Cas9 system,originally derived from the prokaryotic adaptive immune system,has been developed as efficient genome editing tools.It enables precise gene manipulation on chromosomal DNA through the specific b...The CRISPR/Cas9 system,originally derived from the prokaryotic adaptive immune system,has been developed as efficient genome editing tools.It enables precise gene manipulation on chromosomal DNA through the specific binding of programmable sgRNA to target DNA,and the Cas9 protein,which has endonuclease activity,will cut a double strand break at specific locus.However,Cas9 is a foreign protein in mammalian cells,and the potential risks associated with its introduction into mammalian cells are not fully understood.In this study,we performed pull-down and mass spectrometry(MS)analysis of Streptococcus pyogenes Cas9(SpyCas9)interacting proteins in HEK293T cells and showed that the majority of Cas9-associated proteins identified by MS were localized in the nucleolus.Interestingly,we further discovered that the Cas9 protein contains a sequence encoding a nucleolus detention signal(NoDS).Compared with wild-type(WT)Cas9,NoDS-mutated variants of Cas9(mCas9)are less stable,although their gene editing activity is minimally affected.Overexpression of WT Cas9,but not mCas9,causes general effects on transcription and protein translation in the host cell.Overall,identification of NoDS in Cas9 will improve the understanding of Cas9's biological function in vivo,and the removal of NoDS in Cas9 may enhance its safety for future clinical use.展开更多
Purpose:The collaboration relationships between innovation actors at a geographic level may be considered as grouping two separate layers,the domestic and the foreign.At the level of each layer,the relationships and t...Purpose:The collaboration relationships between innovation actors at a geographic level may be considered as grouping two separate layers,the domestic and the foreign.At the level of each layer,the relationships and the actors involved constitute a Triple Helix game.The paper distinguished three levels of analysis:the global grouping together all actors,the domestic grouping together domestic actors,and the foreign related to only actors from partner countries.Design/methodology/approach:Bibliographic records data from the Web of Science for South Korea and West Africa breakdown per innovation actors and distinguishing domestic and international collaboration are analyzed with game theory.The core,the Shapley value,and the nucleolus are computed at the three levels to measure the synergy between actors.Findings:The synergy operates more in South Korea than in West Africa;the government is more present in West Africa than in South Korea;domestic actors create more synergy in South Korea,but foreign more in West Africa;South Korea can consume all the foreign synergy,which is not the case of West Africa.Research limitations:Research data are limited to publication records;techniques and methods used may be extended to other research outputs.Practical implications:West African governments should increase their investment in science,technology,and innovation to benefit more from the synergy their innovation actors contributed at the foreign level.However,the results of the current study may not be sufficient to prove that greater investment will yield benefits from foreign synergies.Originality/value:This paper uses game theory to assess innovation systems by computing the contribution of foreign actors to knowledge production at an area level.It proposes an indicator to this end.展开更多
The dynamic changes of nucleolar ultrastructure in the cell cycle of Physarum polycephalum Schw. were studied by an en bloc silver-staining method. The results showed that the nucleolus was large in size and situated ...The dynamic changes of nucleolar ultrastructure in the cell cycle of Physarum polycephalum Schw. were studied by an en bloc silver-staining method. The results showed that the nucleolus was large in size and situated in the center of the nucleus in late G(2)-phase, and the fibrillar centers, dense fibrillar components and granular components could be observed in the nucleolus. During prophase, the nucleolus moved towards the periphery of the nucleus and in late prophase disintegrated near the nuclear envelope. in metaphase, the disintegrated nucleolar components were dispersed in masses and located at the periphery of the chromosomal region of the nucleus. No specifically silver-stained area and argentophilic protein sheath were observed on the chromosomes, but there were some big dispersed silver particles within the chromosomes. During telophase the nucleolar components moved towards the two poles along with the chromosomes and co-existed with the decondensing chromatin in daughter nuclei. The nucleolar components then gradually converged with one another and separated from the chromatin. A big nucleolus was formed in the nucleus about 120 min after the completion of mitosis.展开更多
The chronological and morphological changes of the nucleus during mouse oocyte maturation and fertilization were systematically studied. Although most oocytes went through GVBD 2-4 hrs after culture, 13.6% remained at...The chronological and morphological changes of the nucleus during mouse oocyte maturation and fertilization were systematically studied. Although most oocytes went through GVBD 2-4 hrs after culture, 13.6% remained at GV stage 8 hrs after culture.TEM observation revealed that nucleoli of oocytes which failed to go through GVBD were composed of fibrillar-granular component,small vacuoles and fibrillar centers or showed small vacuoles on nuclear surface. During GVBD, the nucleoli became smaller and smaller and finally disappeared with the nuclear-associated chromatin dislocated to the periphery. Nuclear membrane with attached chromatin became folded and electronic dense cores appeared in the center of chromatin clumps at the same time.The last event of GVBD was the disruption of nuclear membrane.At the end of the 5th hr after culture, meiosis progressed to prometaphase I.Chromosomes,distributed in the original GV area free of organelles,were surrounded by large quantity of mitochondria and small SER vesicles. At the end of the 12th hr after culture,48. 1% of the oocytes emitted PB1.Decondensing sperm head and early male pronuclcus(mPN)with condensed nucleoli were found 1-2 hrs after insemination.The formation and enlargement of female PN(fPN) occurred a little earlier than that of mPN. 33.3% finished syngamy at 8-9 hrs after insemination.The process of nucleolus formation was reverse to that in GVBD. The oolemma modification caused by cortical reaction could effectively inhibit polyspermy.in contrast,there were sperm binding to the oolemma where CGs failed to be released. In addition, PB2 was emitted 2-5 hrs after insemination. The difference between PB1 and PB2 as well as the abstriction of polar body were also discussed.展开更多
The effects of different concentrations of manganous sulphate on root growth,cell division, chromosome and nucleus morphplogy and nucleoli in root tip-cells of Allium cepa L. were studied. The concentrations of mangan...The effects of different concentrations of manganous sulphate on root growth,cell division, chromosome and nucleus morphplogy and nucleoli in root tip-cells of Allium cepa L. were studied. The concentrations of manganous sulphate used were 10-7,10-6,10-5 ,10-4,10-3, 10-2 and 10-1mol/L. The results indicated that manganese had a stimulatory effect on the root growth at lower concentrations (10-7 to 10-4mol/L). With increasing the concentration of Mn (10-3 to 10-1mol/L) and duration of treatment,manganese inhibited root growth and cell division, and had toxic effects on chromosomes,nuclei and nucleoli. Manganese Could induce mitotic irregularities,comprising c-mitosis,anaphase bridges and chromosome stickiness. The nuclei became irregular in shape and many micronuclei were scattered in the cytoplasm. Some similar silver-stained particulate materials were found scattered in the nucleus in root tip cells. The possible mechanism behind these phenomena is also briefly discussed.展开更多
基金the Shenzhen Science and Technology Research and Development Funds(No.JCYJ20190806155409104)National Natural Science Foundation of China(Nos.52150222,21672130 and 52073163)+1 种基金Guangdong Basic and Applied Basic Research Foundation(No.2019A1515110356)the Qilu Young Scholars Program of Shandong University.
文摘Mitochondrial damage is closely related to the occurrence of many diseases.However,accurate monitoring and reporting of mitochondrial damage are not easy.Here,we developed a small molecule fluorescent probe named CB-Cl,which has splendid spectral properties(large Stokes shift,strong affinity for RNA,etc.)and excellent targeting ability to intracellular mitochondria.After mitochondria were damaged by external stimuli,CB-Cl would light up the nucleolus as a signal reporter.The cascade imaging of mitochondria and nucleolus using CB-Cl can monitor and visualize the mitochondrial status in living cells in real-time.Based on the above advantages,the probe CB-Cl has reference significance for the related research of mitochondrial damage and the prevention and treatment of related diseases.
文摘By using the DNA specific cytochemical staining method (NAMA_Ur) and conventional electron microscopic technique, the authors examined the configuration of intranucleolar DNA in Allium cepa L. cells and found that nucleolar DNA within the fibrillar center (FC) underwent a structural transformation process from condensed to extended state. The authors' observations also displayed a continuous arrangement process of nucleolar DNA, i.e., the extranucleolar DNA entered FC through the nucleolar organizer region (NOR) channel, then extended to the periphery of FC or to the border between FC and dense fibrillar component (DFC), and distributed along the periphery of FC. Thence, by passing through the NOR channel between FCs, the nucleolar DNA continued to transfer to other FCs and arranged in the same above_mentioned forms.
文摘Processing of pre-rRNA is one of the major events taking place In the nucleolus. U3 snoRNA, an rRNA spliceosomal factor, is suggested to be essential in the first cleavage step of the 5' ETS sequence in the processing of pre-rRNA. Identification of U3 in the nucleolus provides a piece of indirect evidence for pre-rRNA processing site and transportation of processing products. In the present study, subnucleolar distribution of U3 snoRNA in the nucleolus of Pisum sativum L. was studied by in situ hybridization with a U3 snoRNA probe. The results showed that the U3 labeling signals were distributed throughout dense fibrillar components (DFCs) and granular components (GCs), while no signal was found in fibrillar centers (FCs). When treated with actinomycine D (AMD), the labeling signals were decreased. Along with the increase of the AMD treatment time, the labeling signals became fewer and they were found in the distal regions of DFC and GC. Our results indicated that pre-rRNA splicing took place in the regions of DFC and GC, and the transportation of pre-rRNA processing products was from the regions around FCs towards the distal regions.
文摘The nucleolus is the most prominent compartment in the nucleus and known as the site for ribosome biogenesis in eucaryotes. In contrast, there is no such equivalent structure for ribosome synthesis in procaryotes. This raises two concerns that how does the nucleolus evolve and that whether the nucleolus remains playing a single role in ribosome biogenesis along the evolution. Increasing data support new nucleolus functions, including signal recognition particle assembly, small RNA modification, telomerase maturation, cell-cycle and aging control, and cell stress sensor. Multiple functions of the nucleolus possibly result from the plurifunctionality of nucleolar proteins, such as nucleolin and Nopp 140. Proteomic analyses of human and Arabidopsis nucleolus lead a remarkable progress in understanding the evolution and new functions of nucleoli. In this review, we present a brief history of nucleolus research and new concepts and unresolved questions. Also, we introduce hepatitis D virus for studying the communication between the nucleolus and other subnuclear compartments, and Caenorhabditis elegans for the role of nucleolus in the development and the epistatic control of nucleologenesis.
基金the National Key R&D Program of China and the Natural Science Foundation of China in the order of 2018YFA0800502,2017YFA0504501,31330050.
文摘The nucleolus,as the‘nucleus of the nucleus’,is a prominent subcellular organelle in a eukaryocyte.The nucleolus serves as the centre for ribosome biogenesis,as well as an important site for cell-cycle regulation,cellular senescence,and stress response.The protein composition of the nucleolus changes dynamically through protein turnover to meet the needs of cellular activities or stress responses.Recent studies have identified a nucleolus-localized protein degradation pathway in zebrafish and humans,namely the Def-CAPN3 pathway,which is essential to ribosome production and cell-cycle progression,by controlling the turnover of multiple substrates(e.g.,ribosomal small-subunit[SSU]processome component Mpp10,transcription factor p53,check-point proteins Chk1 and Wee1).This pathway relies on the Ca2þ-dependent cysteine proteinase CAPN3 and is independent of the ubiquitin-mediated proteasome pathway.CAPN3 is recruited by nucleolar protein Def from cytoplasm to nucleolus,where it proteolyzes its substrates which harbor a CAPN3 recognition-motif.Def depletion leads to the exclusion of CAPN3 and accumulation of p53,Wee1,Chk1,and Mpp10 in the nucleolus that result in cell-cycle arrest and rRNA processing abnormality.Here,we summarize the discovery of the Def-CAPN3 pathway and propose its biological role in cell-cycle control and ribosome biogenesis.
基金The Startup Fund of the Hundred Talents Program of the Chinese Academy of Science (20071010141)National Natural Science Foundation of China(30870120)+1 种基金Open Research Fund Program of the State Key Laboratory of Virology of China (2007003, 2009007)Hubei Province Natural Science Foundation of Innovation Groups Project (2008CDA013)
文摘The nucleolus is a subnuclear structure of eukaryocytes. It was thought that nucleolus only participates in the biogenesis and processing of rRNA. However, more and more evidence shows that it has many other functions, such as tRNA precursor processing, stress sensing and it is also involved in gene silencing, senescence and cell cycle regulation. Here, we summarize the recent understandings about the nucleolar functions, the regulation of nucleolar localization of proteins and the role that the nucleolus plays in virus infection, in which some related studies of Herpes simplex virus type 1 (HSV-1) US11, UL24 and bovine herpesvirus-1 infected cell protein 27 (BICP27) carried out in our lab will also be included.
文摘The effects of different concentrations of cadmium chloride ranging from 0.5 to 20 ppm on the nucleolus in root tip cells of Allium cepa were studied using the silver staining technique. The results indicated that after the treatment with Cd￣(2+),different changes in nucleolar morphology appeared. The nucleolar material was extruded from the nucleus into the cytoplasm,and the nucleoli at mitotic metaphase did not disappear. Apparently,cadmium showed a specific effects on the nucleoli in root tip cells of Allium cepa. The possible mechanism behind this phenomenon is also briefly discussed.
文摘In our conversation with Prof.CHEN Lingling on the"dark mat ter"in the genome,namely the non-coding RNAs,we got to know two classes of long non-coding RNAs(lncRNAs)in close association with Prader-Willi syndrome(PWS).Each with a snoRNA cap on its either end,they fall in the sno-lncRNA family,a group of lncRNAs identified by CHEN’s group in their earlier research.Here we are meeting another member of the family,SLERT,a rescuer
文摘We observed the ultrastructure of nucleolus in rat liver cells by conventional electron microscopy, and employed cytochemistry NAMA-Ur DNA specific stain method to analyze the distribution and position of nucleolar DNA in situ. The results showed that nucleolar DNA of rat liver cells comes from nucleolus-associated chromatin, and continuously extends in the dense fibrillar component (DFC) of nucleolus, localizes at the periphery of fibrillar center (FC) and in DFC. Furthermore, by employing anti-DNA/RNA hybrid antibodies, we directly and selectively labeled transcription sites of rRNA genes and testified that localization of transcription sites not only to DFC but also to the periphery of FC.
基金This work was supported by the National Natural Science Foundation of China (Grant No. 39770367).
文摘The location and configuration of nucleolar DNA have not been determined for a long time. In this paper, we have observed the nucleolar infrastructure and the character of nucleolar DNA in Allium cepa cells by conventional electron microscopy and the cytochemical NAMA-Ur DNA specific staining method. Furthermore, we have properly improved the NAMA-Ur method so as to analyze the location and configuration of nucleolar DNA in situ. Our results indicated that the nucleolar DNA in Allium cepa cells is mainly located at the border between fibrillar centers and dense fibribllar component, especially distributed in the configuration of encircling the fibrillar centers.
基金part of the Dutch Research Council(NWO)Building Blocks of Lite programme(grant number 737.016.007).
文摘Microalgae are used in food and feed,and they are considered a potential feedstock for sustainably pro-duced chemicals and biofuel.However,production of microalgal-derived chemicals is not yet economically feasible.Genetic engineering could bridge the gap to industrial application and facilitate the production of novel products from microalgae.Here,we report the discovery of a novel gene expression system in the oleaginous microalga Nannochloropsis that exploits the highly efficient transcriptional activity of RNA po-lymerase I and an internal ribosome entry site for translation.We identified the nucleolus as a genomic safe harbor for Pol I transcription and used it to construct transformant strains with consistently strong trans-gene expression.The new expression system provides an outstanding tool for genetic and metabolic en-gineering of microalgae and thus will probably make substantial contributions to microalgal research.
文摘Nucleolus organizer regions (Nor loci) of a range of Chinese wheat landraces and cultivars (Triticum aestivum L. em Thell.) were analysed using genomic DNA extracted from leaves. Only two allelic variants of the Nor-B1 locus were found on chromosome 1B (Nor-B1a and Nor-B1g), while Nor-B1g was probably introduced from North America in the early 1960s. The even more recent introduction of the rye allele Nor-R1 in the early 1980s was also revealed. Eight allelic variants of the Nor-B2 locus on chromosome 6B (Nor-B2a, b, d, f, h, o, p and s) were identified. A Chinese origin for the a, d, f, o, p and s alkies is evident although the d allele was successfully introduced into Australian wheats in the early 1900s. Nor-B2h and Nor-B2b are again very recent introductions into Chinese wheat breeding programs, the former from CIMMYT wheats and the latter in association with the introduction of the 1RS/1BL translocation from Europe. On the basis of the presence of different combinations of Nor-B1 and Nor-B2 alleles, the Chinese wheat cultivars can be divided into eight phenotypic groups.
文摘Nucleolus is the most prominent subnuclear structure, which performs a wide variety of functions in the eukaryotic cellular processes. In order to understand the structural and functional role of the nucleoli in bovine cells, we analyzed the proteomic composition of the bovine nucleoli. The nucleoli were isolated from Madin Darby bovine kidney cells and subjected to proteomic analysis by LC-MS/MS after fractionation by SDS-PAGE and strong cation exchange chromatography. Analysis of the data using the Mascot database search and the GPM database search identified 311 proteins in the bovine nucleoli, which contained 22 proteins previously not identified in the proteomic analysis of human nucleoli. Analysis of the identified proteins using the GoMiner software suggested that the bovine nucleoli contained proteins involved in ribosomal biogenesis, cell cycle control, transcriptional, translational and post-translational regulation, transport, and structural organization.
文摘Our conventional EM observations indicate that the nucleolus of Allium cepa is composedof the fibrillar centre (FC), fibrillar component (F) and granular region (G). FC is an elec-tron-lucid zone which contains condensed or loosened chromatin. F is a circular, highly elec-tron-dense component which surrounds FC and consists of compact fibrils. EM autoradiographic(EM ARG) studies with ~3H- UdR reveal that the synthesis of ribosomal RNA (rRNA) takesplace in F. G is situated around F and composed of granules about 25 to 30 nm in diameter.EM ARG studies with ~3H- TdR demonstrate that silver grains are predominantly located in Gof the nucleolar periphery and the region of the nucleolus-associated chromatin. In addition,~3H- TdR labels are also present, but with a much lower frequency, in FC. F and G of the nu-cleolar interior. According to the distribution and sphere of silver grains in different compo-nents of the nucleolus, we suggest that the replication sites of ribosomal DNA (rDNA) aremainly located in G of the nucleolar periphery and the region of the nucleolus-associated chro-matin. Our observations also suggest that some segments of rDNA initiate their replication inFC and then move to the nucleolar periphery.
基金This work was supported by the National Key Research and Development Program of China(No.2018YFC0310900)the National Natural Science Foundation of China grants(No.31270830,21572038,21877016 to Y.D,No.31671386 to Q.H,81972621 to W.J)+1 种基金the Development Fund for Shanghai Talents,Fund of State Key Laboratory of Bioorganic and Natural Products Chemistry,Fund of State Key Laboratory of Drug Research,Chinese Academy of Science(No.SIMM1601KF-08)Fudan University Graduate Student Research Grant.
文摘The CRISPR/Cas9 system,originally derived from the prokaryotic adaptive immune system,has been developed as efficient genome editing tools.It enables precise gene manipulation on chromosomal DNA through the specific binding of programmable sgRNA to target DNA,and the Cas9 protein,which has endonuclease activity,will cut a double strand break at specific locus.However,Cas9 is a foreign protein in mammalian cells,and the potential risks associated with its introduction into mammalian cells are not fully understood.In this study,we performed pull-down and mass spectrometry(MS)analysis of Streptococcus pyogenes Cas9(SpyCas9)interacting proteins in HEK293T cells and showed that the majority of Cas9-associated proteins identified by MS were localized in the nucleolus.Interestingly,we further discovered that the Cas9 protein contains a sequence encoding a nucleolus detention signal(NoDS).Compared with wild-type(WT)Cas9,NoDS-mutated variants of Cas9(mCas9)are less stable,although their gene editing activity is minimally affected.Overexpression of WT Cas9,but not mCas9,causes general effects on transcription and protein translation in the host cell.Overall,identification of NoDS in Cas9 will improve the understanding of Cas9's biological function in vivo,and the removal of NoDS in Cas9 may enhance its safety for future clinical use.
文摘Purpose:The collaboration relationships between innovation actors at a geographic level may be considered as grouping two separate layers,the domestic and the foreign.At the level of each layer,the relationships and the actors involved constitute a Triple Helix game.The paper distinguished three levels of analysis:the global grouping together all actors,the domestic grouping together domestic actors,and the foreign related to only actors from partner countries.Design/methodology/approach:Bibliographic records data from the Web of Science for South Korea and West Africa breakdown per innovation actors and distinguishing domestic and international collaboration are analyzed with game theory.The core,the Shapley value,and the nucleolus are computed at the three levels to measure the synergy between actors.Findings:The synergy operates more in South Korea than in West Africa;the government is more present in West Africa than in South Korea;domestic actors create more synergy in South Korea,but foreign more in West Africa;South Korea can consume all the foreign synergy,which is not the case of West Africa.Research limitations:Research data are limited to publication records;techniques and methods used may be extended to other research outputs.Practical implications:West African governments should increase their investment in science,technology,and innovation to benefit more from the synergy their innovation actors contributed at the foreign level.However,the results of the current study may not be sufficient to prove that greater investment will yield benefits from foreign synergies.Originality/value:This paper uses game theory to assess innovation systems by computing the contribution of foreign actors to knowledge production at an area level.It proposes an indicator to this end.
文摘The dynamic changes of nucleolar ultrastructure in the cell cycle of Physarum polycephalum Schw. were studied by an en bloc silver-staining method. The results showed that the nucleolus was large in size and situated in the center of the nucleus in late G(2)-phase, and the fibrillar centers, dense fibrillar components and granular components could be observed in the nucleolus. During prophase, the nucleolus moved towards the periphery of the nucleus and in late prophase disintegrated near the nuclear envelope. in metaphase, the disintegrated nucleolar components were dispersed in masses and located at the periphery of the chromosomal region of the nucleus. No specifically silver-stained area and argentophilic protein sheath were observed on the chromosomes, but there were some big dispersed silver particles within the chromosomes. During telophase the nucleolar components moved towards the two poles along with the chromosomes and co-existed with the decondensing chromatin in daughter nuclei. The nucleolar components then gradually converged with one another and separated from the chromatin. A big nucleolus was formed in the nucleus about 120 min after the completion of mitosis.
文摘The chronological and morphological changes of the nucleus during mouse oocyte maturation and fertilization were systematically studied. Although most oocytes went through GVBD 2-4 hrs after culture, 13.6% remained at GV stage 8 hrs after culture.TEM observation revealed that nucleoli of oocytes which failed to go through GVBD were composed of fibrillar-granular component,small vacuoles and fibrillar centers or showed small vacuoles on nuclear surface. During GVBD, the nucleoli became smaller and smaller and finally disappeared with the nuclear-associated chromatin dislocated to the periphery. Nuclear membrane with attached chromatin became folded and electronic dense cores appeared in the center of chromatin clumps at the same time.The last event of GVBD was the disruption of nuclear membrane.At the end of the 5th hr after culture, meiosis progressed to prometaphase I.Chromosomes,distributed in the original GV area free of organelles,were surrounded by large quantity of mitochondria and small SER vesicles. At the end of the 12th hr after culture,48. 1% of the oocytes emitted PB1.Decondensing sperm head and early male pronuclcus(mPN)with condensed nucleoli were found 1-2 hrs after insemination.The formation and enlargement of female PN(fPN) occurred a little earlier than that of mPN. 33.3% finished syngamy at 8-9 hrs after insemination.The process of nucleolus formation was reverse to that in GVBD. The oolemma modification caused by cortical reaction could effectively inhibit polyspermy.in contrast,there were sperm binding to the oolemma where CGs failed to be released. In addition, PB2 was emitted 2-5 hrs after insemination. The difference between PB1 and PB2 as well as the abstriction of polar body were also discussed.
文摘The effects of different concentrations of manganous sulphate on root growth,cell division, chromosome and nucleus morphplogy and nucleoli in root tip-cells of Allium cepa L. were studied. The concentrations of manganous sulphate used were 10-7,10-6,10-5 ,10-4,10-3, 10-2 and 10-1mol/L. The results indicated that manganese had a stimulatory effect on the root growth at lower concentrations (10-7 to 10-4mol/L). With increasing the concentration of Mn (10-3 to 10-1mol/L) and duration of treatment,manganese inhibited root growth and cell division, and had toxic effects on chromosomes,nuclei and nucleoli. Manganese Could induce mitotic irregularities,comprising c-mitosis,anaphase bridges and chromosome stickiness. The nuclei became irregular in shape and many micronuclei were scattered in the cytoplasm. Some similar silver-stained particulate materials were found scattered in the nucleus in root tip cells. The possible mechanism behind these phenomena is also briefly discussed.
