AT-hook是一类新的DNA结合蛋白基序,与其他功能已知的DNA结合蛋白基序不同。AT-hook蛋白具有AT-hook基序和PPC(plants and prokaryotes conserved domain,DUF296)两个特殊功能域。AT-hook广泛存在于不同物种的DNA结合蛋白中,在植物生长...AT-hook是一类新的DNA结合蛋白基序,与其他功能已知的DNA结合蛋白基序不同。AT-hook蛋白具有AT-hook基序和PPC(plants and prokaryotes conserved domain,DUF296)两个特殊功能域。AT-hook广泛存在于不同物种的DNA结合蛋白中,在植物生长发育、器官构建、逆境胁迫和激素信号应答中发挥重要的调节作用;对基因克隆、细胞间特异性结合、染色体结构调节以及转录因子调节具有重要的调控作用。通过调节AT-hook蛋白进而改变生物某些不理想的生理生化调控通路,提高生物某些优良性状具有重要研究价值及意义。本综述主要从AT-hook蛋白的结构特征、分类及其依据、功能调节机制、生物学功能以及研究价值等方面进行相关阐述及总结。展开更多
Class Ⅲ peroxidases(CⅢ Prxs) play critical roles in plant immunity by scavenging reactive oxygen species(ROS). However, the functions of CⅢ Prxs in rice(Oryza sativa L.) immunity are largely unexplored. Here, we re...Class Ⅲ peroxidases(CⅢ Prxs) play critical roles in plant immunity by scavenging reactive oxygen species(ROS). However, the functions of CⅢ Prxs in rice(Oryza sativa L.) immunity are largely unexplored. Here, we report a Prx precursor, OsPrx30,that is responsive to the bacterial blight Xanthomonas oryzae pv. oryzae(Xoo). OsPrx30 was primarily expressed in rice roots, leaves, and stems,and its protein product was mainly localized at the endoplasmic reticulum. Overexpression of OsPrx30 enhanced the plant’s susceptibility to Xoo by maintaining a high level of peroxidase(POD) activity and reducing the content of H2O2, whereas depletion of OsPrx30 had the opposite effects. Furthermore, we identified an AT-hook transcription factor, Os ATH1, that is specifically bound to the OsPrx30 promoter. As observed in plants overexpressing OsPrx30, depletion of Os ATH1 enhanced susceptibility to Xoo. Finally, we demonstrated that depletion of Os ATH1 increased histone H3 acetylation at the AT-rich region of the OsPrx30 promoter.Taken together, these results reveal a mechanism underlying the POD-induced natural resistance to bacterial diseases and suggest a model for transcription regulation of Prx genes in rice.展开更多
Hypocotyl length is regarded to be a crucial seedling trait,influencing many subsequent plant development processes.However,little is known about this trait in Brassica campestris syn.Brasscia rapa.Here,we performed a...Hypocotyl length is regarded to be a crucial seedling trait,influencing many subsequent plant development processes.However,little is known about this trait in Brassica campestris syn.Brasscia rapa.Here,we performed a comparative observation on the early hypocotyl development between two cultivars,‘SZQ’belonging to pak-choi(B.campestris ssp.chinensis var.communis)with longer hypocotyls,and‘WTC’belonging to Tacai(B.campestris L.ssp.chinensis var.rosularis)with shortter hypocotyls,and found that the difference in auxin biosynthesis might contribute to the varied hypocotyl phenotype between these two cultivars.By applying GWAS analysis using a total of 226 B.campestris accessions,we identified that the AT-Hook motif nuclear localized(AHL)gene BcAHL24-MF1 contributed to the natural variation in hypocotyl length.Functional variation of BcAHL24-MF1 was attributed to four haplotypes featuring four SNPs within the promoter region,of which Hap I accumulated more transcripts with shorter hypocotyls.Constitutive overexpression of BcAHL24-MF1 in B.campestris caused decreased hypocotyl length under light circumstances and even constant darkness,as BcAHL24-MF1 repressed the PIFmediated transcriptional activation of auxin biosynthesis genes BcYUC6-MF2 and BcYUC8-LF.Our research uncovered the important role of BcAHL24-MF1 in regulating light-triggered inhibition of hypocotyl elongation,therefore presenting a valuable genetic target for crop breeding.展开更多
Background:Skin innervation is very important for normal wound healing,and receptor activity-modifying protein 1(RAMP1)has been reported to modulate calcitonin gene-related peptide(CGRP)receptor function and thus be a...Background:Skin innervation is very important for normal wound healing,and receptor activity-modifying protein 1(RAMP1)has been reported to modulate calcitonin gene-related peptide(CGRP)receptor function and thus be a potential treatment target.This study aimed to elucidate the intricate regulatory effect of RAMP1 on skin fibroblast function,thereby addressing the existing knowledge gap in this area.Methods:Immunohistochemical staining and immunofluorescence(IF)staining were used to measure the dynamic changes in the expression of RAMP1 and α-smooth muscle actin(α-SMA)in skin wound tissue in mice.Mouse skin fibroblasts(MSFs)stably transfected with Tet-on-Flag-RAMP1 overexpression(OE)and Tet-on-Flag control(Ctrl)lentiviruses were constructed for in vitro experiments.High mobility group AT-hook 1(HMGA1)plasmids and α-SMA plasmids were used to overexpress HMGA1 and α-SMA,respectively.Anα-SMA siRNA was used to silence α-SMA.Quantitative real-time polymerase chain reaction(qPCR),western blot and IF staining analyses were used to determine the mRNA and protein levels in the cells in different groups.A scratch wound healing assay was used to evaluate the cell migration ability of different groups.Cleavage under targets and release using nuclease(CUT&RUN)assays and dual-luciferase reporter assays were used to predict and verify the interaction between HMGA1 and the α-SMA promoter.Results:RAMP1 and α-SMA protein expression levels in the dermis changed dynamically and were negatively correlated during dorsal skin wound healing in mice.RAMP1 OE in vitro inhibited the differentiation and promoted the migration of MSFs by decreasingα-SMA expression via the suppression of HMGA1,which was shown for the first time to bind to theα-SMA promoter and increase α-SMA transcription.RAMP1 OE also modulated extracellular matrix(ECM)synthesis and remodeling by promoting collagen Ⅲ and MMP9 expression and decreasing collagen Ⅰ,MMP2,and tissue inhibitor of metalloproteinases 1 expression.Conclusions:Our findings suggest that RAMP1 OE decreases differentiation and promotes migration in MSFs by downregulating α-SMA expression via the suppression of HMGA1 and modulates ECM synthesis and remodeling,revealing a novel mechanism regulating α-SMA transcription,providing new insights into the RAMP1-mediated regulation of fibroblast function,and identifying effective nerve-related targets for skin wound repair.展开更多
Objective:To analyze effects of high mobility group AT-hook 2(HMGA2)on malignant degree,invasion,metastasis,proliferation and cellular morphology of ovarian cancer cells.Methods:Three methods were applied to observe t...Objective:To analyze effects of high mobility group AT-hook 2(HMGA2)on malignant degree,invasion,metastasis,proliferation and cellular morphology of ovarian cancer cells.Methods:Three methods were applied to observe the effect on HMGA2 expression in ovarian cancer cells and ovarian epithelial cells.Results:After the application of siRNA-HMGA2,number of T29A2-cell clones was decreased,there was significant difference compared with the negative control Block-iT.After application of let-7c,number of T29A2+cell clones was decreased significantly,however,after the application of Anti-let-7,the number of clones restored,and there was no significant difference compared with the negative control group.After interference,the number of T29A2-cells which passed through Matrigel polycarbonate membrane were significantly lower than the negative control group.After the treatment of siRNA-HMGA2,let-7c and sh-HMGA2respectively,growth and proliferation of T29A2-,T29A2+and SKOV3 were slower,and the phenomenon was most obvious in SKOV3.Stable interference of HMGA2 induced mesenchymalepithelial changes in the morphology of SKOV3-sh-HMGA2.Conclusions:HMGA2 can promote malignant transformation of ovarian cancer cells,enhance cell invasion and metastasis,and promote cell growth and proliferation of ovarian cancer cells,which can cause ovarian cancer to progress rapidly and affect the quality of life.展开更多
文摘AT-hook是一类新的DNA结合蛋白基序,与其他功能已知的DNA结合蛋白基序不同。AT-hook蛋白具有AT-hook基序和PPC(plants and prokaryotes conserved domain,DUF296)两个特殊功能域。AT-hook广泛存在于不同物种的DNA结合蛋白中,在植物生长发育、器官构建、逆境胁迫和激素信号应答中发挥重要的调节作用;对基因克隆、细胞间特异性结合、染色体结构调节以及转录因子调节具有重要的调控作用。通过调节AT-hook蛋白进而改变生物某些不理想的生理生化调控通路,提高生物某些优良性状具有重要研究价值及意义。本综述主要从AT-hook蛋白的结构特征、分类及其依据、功能调节机制、生物学功能以及研究价值等方面进行相关阐述及总结。
基金supported by the Natural Science Foundation of Guangdong Province of China (2018A0303130340)the Guangdong Basic and Applied Basic Research Foundation (2020A1515010021)+1 种基金the Research and Development Program in key areas of Guangdong Province (2018B020206002)the National Natural Science Foundation of China (31401722)。
文摘Class Ⅲ peroxidases(CⅢ Prxs) play critical roles in plant immunity by scavenging reactive oxygen species(ROS). However, the functions of CⅢ Prxs in rice(Oryza sativa L.) immunity are largely unexplored. Here, we report a Prx precursor, OsPrx30,that is responsive to the bacterial blight Xanthomonas oryzae pv. oryzae(Xoo). OsPrx30 was primarily expressed in rice roots, leaves, and stems,and its protein product was mainly localized at the endoplasmic reticulum. Overexpression of OsPrx30 enhanced the plant’s susceptibility to Xoo by maintaining a high level of peroxidase(POD) activity and reducing the content of H2O2, whereas depletion of OsPrx30 had the opposite effects. Furthermore, we identified an AT-hook transcription factor, Os ATH1, that is specifically bound to the OsPrx30 promoter. As observed in plants overexpressing OsPrx30, depletion of Os ATH1 enhanced susceptibility to Xoo. Finally, we demonstrated that depletion of Os ATH1 increased histone H3 acetylation at the AT-rich region of the OsPrx30 promoter.Taken together, these results reveal a mechanism underlying the POD-induced natural resistance to bacterial diseases and suggest a model for transcription regulation of Prx genes in rice.
基金supported by grants from the Key R and D Program of Zhejiang(Grant Nos.2022C02032 and 2022C02030)the SanNong JiuFang Science and Technology Cooperation Project of Zhejiang Province(Grant No.2023SNJF008)+1 种基金the Grand Science and Technology Special Project of Zhejiang Province(Grant No.2021C02065)the Science and Technology Plan Project of Jiaxing(Grant No.2023AZ11002).
文摘Hypocotyl length is regarded to be a crucial seedling trait,influencing many subsequent plant development processes.However,little is known about this trait in Brassica campestris syn.Brasscia rapa.Here,we performed a comparative observation on the early hypocotyl development between two cultivars,‘SZQ’belonging to pak-choi(B.campestris ssp.chinensis var.communis)with longer hypocotyls,and‘WTC’belonging to Tacai(B.campestris L.ssp.chinensis var.rosularis)with shortter hypocotyls,and found that the difference in auxin biosynthesis might contribute to the varied hypocotyl phenotype between these two cultivars.By applying GWAS analysis using a total of 226 B.campestris accessions,we identified that the AT-Hook motif nuclear localized(AHL)gene BcAHL24-MF1 contributed to the natural variation in hypocotyl length.Functional variation of BcAHL24-MF1 was attributed to four haplotypes featuring four SNPs within the promoter region,of which Hap I accumulated more transcripts with shorter hypocotyls.Constitutive overexpression of BcAHL24-MF1 in B.campestris caused decreased hypocotyl length under light circumstances and even constant darkness,as BcAHL24-MF1 repressed the PIFmediated transcriptional activation of auxin biosynthesis genes BcYUC6-MF2 and BcYUC8-LF.Our research uncovered the important role of BcAHL24-MF1 in regulating light-triggered inhibition of hypocotyl elongation,therefore presenting a valuable genetic target for crop breeding.
基金supported by the National Natural Science Foundation of China(81972947 to YBW)the Natural Science Foundation of Shandong Province of China(Major Basic Research Program)(ZR2019ZD38 to YBW)+2 种基金the Key Research and Development Program of Shandong Province of China(No.2019GSF108128 to YBW)the Academic Promotion Programme of Shandong First Medical University(2019LJ005 to YBW)the Jinan Clinical Research Center for Tissue Engineering Skin Regeneration and Wound Repair.
文摘Background:Skin innervation is very important for normal wound healing,and receptor activity-modifying protein 1(RAMP1)has been reported to modulate calcitonin gene-related peptide(CGRP)receptor function and thus be a potential treatment target.This study aimed to elucidate the intricate regulatory effect of RAMP1 on skin fibroblast function,thereby addressing the existing knowledge gap in this area.Methods:Immunohistochemical staining and immunofluorescence(IF)staining were used to measure the dynamic changes in the expression of RAMP1 and α-smooth muscle actin(α-SMA)in skin wound tissue in mice.Mouse skin fibroblasts(MSFs)stably transfected with Tet-on-Flag-RAMP1 overexpression(OE)and Tet-on-Flag control(Ctrl)lentiviruses were constructed for in vitro experiments.High mobility group AT-hook 1(HMGA1)plasmids and α-SMA plasmids were used to overexpress HMGA1 and α-SMA,respectively.Anα-SMA siRNA was used to silence α-SMA.Quantitative real-time polymerase chain reaction(qPCR),western blot and IF staining analyses were used to determine the mRNA and protein levels in the cells in different groups.A scratch wound healing assay was used to evaluate the cell migration ability of different groups.Cleavage under targets and release using nuclease(CUT&RUN)assays and dual-luciferase reporter assays were used to predict and verify the interaction between HMGA1 and the α-SMA promoter.Results:RAMP1 and α-SMA protein expression levels in the dermis changed dynamically and were negatively correlated during dorsal skin wound healing in mice.RAMP1 OE in vitro inhibited the differentiation and promoted the migration of MSFs by decreasingα-SMA expression via the suppression of HMGA1,which was shown for the first time to bind to theα-SMA promoter and increase α-SMA transcription.RAMP1 OE also modulated extracellular matrix(ECM)synthesis and remodeling by promoting collagen Ⅲ and MMP9 expression and decreasing collagen Ⅰ,MMP2,and tissue inhibitor of metalloproteinases 1 expression.Conclusions:Our findings suggest that RAMP1 OE decreases differentiation and promotes migration in MSFs by downregulating α-SMA expression via the suppression of HMGA1 and modulates ECM synthesis and remodeling,revealing a novel mechanism regulating α-SMA transcription,providing new insights into the RAMP1-mediated regulation of fibroblast function,and identifying effective nerve-related targets for skin wound repair.
基金广东省基础与应用基础研究重大项目(2021B0301030004)云南省William John Lucas院士工作站(202105AF150028)+2 种基金国家自然科学基金项目“二倍体栽培种马铃薯RH中自交亲和基因的图位克隆”(32002032)云南省基础研究专项面上项目“薯肉龙葵素含量负调控因子StERF9基因的克隆和功能解析”(202201AT070037)昭通市千和农业科技开发有限公司“尚轶专家工作站”项目资助。
基金supported by National Natural Science Foundation of China(6821183728)
文摘Objective:To analyze effects of high mobility group AT-hook 2(HMGA2)on malignant degree,invasion,metastasis,proliferation and cellular morphology of ovarian cancer cells.Methods:Three methods were applied to observe the effect on HMGA2 expression in ovarian cancer cells and ovarian epithelial cells.Results:After the application of siRNA-HMGA2,number of T29A2-cell clones was decreased,there was significant difference compared with the negative control Block-iT.After application of let-7c,number of T29A2+cell clones was decreased significantly,however,after the application of Anti-let-7,the number of clones restored,and there was no significant difference compared with the negative control group.After interference,the number of T29A2-cells which passed through Matrigel polycarbonate membrane were significantly lower than the negative control group.After the treatment of siRNA-HMGA2,let-7c and sh-HMGA2respectively,growth and proliferation of T29A2-,T29A2+and SKOV3 were slower,and the phenomenon was most obvious in SKOV3.Stable interference of HMGA2 induced mesenchymalepithelial changes in the morphology of SKOV3-sh-HMGA2.Conclusions:HMGA2 can promote malignant transformation of ovarian cancer cells,enhance cell invasion and metastasis,and promote cell growth and proliferation of ovarian cancer cells,which can cause ovarian cancer to progress rapidly and affect the quality of life.
