During seed germination,the cotton chaperone protein HSP24.7 regulates the release,from the mitochondrial electron transport chain,of reactive oxygen species(ROS),a stimulative signal regulating germination.The functi...During seed germination,the cotton chaperone protein HSP24.7 regulates the release,from the mitochondrial electron transport chain,of reactive oxygen species(ROS),a stimulative signal regulating germination.The function of HSP24.7 during vegetative stages remains largely unknown.Here we propose that suppression of Gh HSP24.7 in cotton seedlings increases tolerance to heat and drought stress.Elevation of Gh HSP24.7 was found to be positively associated with endogenous levels of ROS.We identified a new client protein of Gh HSP24.7,cotton lysine deacetylase(Gh HDA14),which is involved in mitochondrial protein modification.Elevated levels of Gh HSP24.7 suppressed deacetylase activity in mitochondria,leading to increased acetylation of mitochondrial proteins enriched in the subunit of Ftype ATPase,V-type ATPase,and cytochrome C reductase,ultimately reducing leaf ATP content.Consequently,in combination with altered ROS content,Gh HSP24.7 transgenic lines were unable to coordinate stomatal closure under stress.The regulation circuit composed of Gh HSP24.7 and Gh HDA14 represents a post-translation level mechanism in plant abiotic stress responses that integrates the regulation of ROS and ATP.展开更多
AIM: To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells(EPCs) in a mouse model of laser-induced retinal injury.METHODS: EPCs were isolated from ...AIM: To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells(EPCs) in a mouse model of laser-induced retinal injury.METHODS: EPCs were isolated from human umbilical cord blood mononuclear cells and labeled with three different fluorescent probes: 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester(CFSE), 1,1′-dilinoleyl-3,3,3′,3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein(Di I-Ac LDL), and green fluorescent protein(GFP). The fluorescent intensity of EPCs was examined by confocal microscopy. Survival rate of labeled EPCs was calculated with trypan blue staining, and their adhesive capability was assessed. A mouse model of retinal injury was induced by laser, and EPCs were injected into the vitreous cavity. Frozen section and fluorescein angiography on flat-mounted retinal samples was employed to track the labeled EPCs in vivo.RESULTS: EPCs labeled with CFSE and Di I-Ac LDL exhibited an intense green and red fluorescence at the beginning; the fluorescence intensity decreased gradually to 20.23% and 49.99% respectively, after 28 d. On the contrary, the florescent intensity of GFP-labeled EPCs increased in a time-dependent manner. All labeled EPCs showed normal morphology and no significant change in survival and adhesive capability. In the mouse model, transplantation of EPCs showed a protective effect against retinal injury. EPCs labeled with CFSE and Di I-Ac LDL were successfully tracked in mice during the development of retinal injury and repair; however, GFP-labeled EPCs were not detected in the laser-injured mouse retina.CONCLUSION: The three fluorescent markers used in this study have their own set of advantages and disadvantages. CFSE and Di I-Ac LDL are suitable for short-term EPClabeling, while GFP should be used for long-term labeling. The choice of fluorescent markers should be guided by the purpose of the study.展开更多
Post-translational modifications(PTMs) occurring at protein lysine residues,or protein lysine modifications(PLMs),play critical roles in regulating biological processes.Due to the explosive expansion of the amount...Post-translational modifications(PTMs) occurring at protein lysine residues,or protein lysine modifications(PLMs),play critical roles in regulating biological processes.Due to the explosive expansion of the amount of PLM substrates and the discovery of novel PLM types,here we greatly updated our previous studies,and presented a much more integrative resource of protein lysine modification database(PLMD).In PLMD,we totally collected and integrated 284,780 modification events in 53,501 proteins across 176 eukaryotes and prokaryotes for up to 20 types of PLMs,including ubiquitination, acetylation, sumoylation, methylation ,succinylation,malonylation,glutarylation,giycation,formylation,hydroxylation,butyrylation,propionylation,crotonylation,pupylation,neddylation,2-hydroxyisobutyrylation,phosphoglycerylation,carboxylation,lipoylation and biotinylation.Using the data set,a motif-based analysis was performed for each PLM type,and the results demonstrated that different PLM types preferentially recognize distinct sequence motifs for the modifications.Moreover,various PLMs synergistically orchestrate specific cellular biological processes by mutual crosstalks with each other,and we totally found 65,297 PLM events involved in 90 types of PLM co-occurrences on the same lysine residues.Finally,various options were provided for accessing the data,while original references and other annotations were also present for each PLM substrate.Taken together,we anticipated the PLMD database can serve as a useful resource for further researches of PLMs.PLMD 3.0 was implemented in PHP + MySQL and freely available at http://plmd.biocuckoo.org.展开更多
基金financially supported by the National Natural Science Foundation of China(NSFC 31901430,31971985)Leading Innovative and Entrepreneur Team Introduction Program of Zhejiang(2019R01002)+1 种基金Fundamental Research Funds for the Central UniversitiesJiangsu Collaborative Innovation Center for Modern Crop Production(JCIC-MCP)。
文摘During seed germination,the cotton chaperone protein HSP24.7 regulates the release,from the mitochondrial electron transport chain,of reactive oxygen species(ROS),a stimulative signal regulating germination.The function of HSP24.7 during vegetative stages remains largely unknown.Here we propose that suppression of Gh HSP24.7 in cotton seedlings increases tolerance to heat and drought stress.Elevation of Gh HSP24.7 was found to be positively associated with endogenous levels of ROS.We identified a new client protein of Gh HSP24.7,cotton lysine deacetylase(Gh HDA14),which is involved in mitochondrial protein modification.Elevated levels of Gh HSP24.7 suppressed deacetylase activity in mitochondria,leading to increased acetylation of mitochondrial proteins enriched in the subunit of Ftype ATPase,V-type ATPase,and cytochrome C reductase,ultimately reducing leaf ATP content.Consequently,in combination with altered ROS content,Gh HSP24.7 transgenic lines were unable to coordinate stomatal closure under stress.The regulation circuit composed of Gh HSP24.7 and Gh HDA14 represents a post-translation level mechanism in plant abiotic stress responses that integrates the regulation of ROS and ATP.
基金Supported by the National Natural Science Foundation of China(No.81400403)the International Science and Technology Cooperation Program of Jilin Province(No.20110733)the Technology Program of Soochow City(No.SYS201375)
文摘AIM: To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells(EPCs) in a mouse model of laser-induced retinal injury.METHODS: EPCs were isolated from human umbilical cord blood mononuclear cells and labeled with three different fluorescent probes: 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester(CFSE), 1,1′-dilinoleyl-3,3,3′,3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein(Di I-Ac LDL), and green fluorescent protein(GFP). The fluorescent intensity of EPCs was examined by confocal microscopy. Survival rate of labeled EPCs was calculated with trypan blue staining, and their adhesive capability was assessed. A mouse model of retinal injury was induced by laser, and EPCs were injected into the vitreous cavity. Frozen section and fluorescein angiography on flat-mounted retinal samples was employed to track the labeled EPCs in vivo.RESULTS: EPCs labeled with CFSE and Di I-Ac LDL exhibited an intense green and red fluorescence at the beginning; the fluorescence intensity decreased gradually to 20.23% and 49.99% respectively, after 28 d. On the contrary, the florescent intensity of GFP-labeled EPCs increased in a time-dependent manner. All labeled EPCs showed normal morphology and no significant change in survival and adhesive capability. In the mouse model, transplantation of EPCs showed a protective effect against retinal injury. EPCs labeled with CFSE and Di I-Ac LDL were successfully tracked in mice during the development of retinal injury and repair; however, GFP-labeled EPCs were not detected in the laser-injured mouse retina.CONCLUSION: The three fluorescent markers used in this study have their own set of advantages and disadvantages. CFSE and Di I-Ac LDL are suitable for short-term EPClabeling, while GFP should be used for long-term labeling. The choice of fluorescent markers should be guided by the purpose of the study.
基金supported by grants from the National Basic Research Program(973 projectNo.2013CB933900)+1 种基金the Natural Science Foundation of China(Nos.31671360 andJ1103514)the International Science & Technology Cooperation Program of China (No.2014DFB30020)
文摘Post-translational modifications(PTMs) occurring at protein lysine residues,or protein lysine modifications(PLMs),play critical roles in regulating biological processes.Due to the explosive expansion of the amount of PLM substrates and the discovery of novel PLM types,here we greatly updated our previous studies,and presented a much more integrative resource of protein lysine modification database(PLMD).In PLMD,we totally collected and integrated 284,780 modification events in 53,501 proteins across 176 eukaryotes and prokaryotes for up to 20 types of PLMs,including ubiquitination, acetylation, sumoylation, methylation ,succinylation,malonylation,glutarylation,giycation,formylation,hydroxylation,butyrylation,propionylation,crotonylation,pupylation,neddylation,2-hydroxyisobutyrylation,phosphoglycerylation,carboxylation,lipoylation and biotinylation.Using the data set,a motif-based analysis was performed for each PLM type,and the results demonstrated that different PLM types preferentially recognize distinct sequence motifs for the modifications.Moreover,various PLMs synergistically orchestrate specific cellular biological processes by mutual crosstalks with each other,and we totally found 65,297 PLM events involved in 90 types of PLM co-occurrences on the same lysine residues.Finally,various options were provided for accessing the data,while original references and other annotations were also present for each PLM substrate.Taken together,we anticipated the PLMD database can serve as a useful resource for further researches of PLMs.PLMD 3.0 was implemented in PHP + MySQL and freely available at http://plmd.biocuckoo.org.