BACKGROUND The hypoxic environment during bone healing is important in regulating the differentiation of periosteal stem cells(PSCs)into osteoblasts or chondrocytes;however,the underlying mechanisms remain unclear.AIM...BACKGROUND The hypoxic environment during bone healing is important in regulating the differentiation of periosteal stem cells(PSCs)into osteoblasts or chondrocytes;however,the underlying mechanisms remain unclear.AIM To determine the effect of hypoxia on PSCs,and the expression of microRNA-584-5p(miR-584-5p)and RUNX family transcription factor 2(RUNX2)in PSCs was modulated to explore the impact of the miR-584-5p/RUNX2 axis on hypoxiainduced osteogenic differentiation of PSCs.METHODS In this study,we isolated primary mouse PSCs and stimulated them with hypoxia,and the characteristics and functional genes related to PSC osteogenic differentiation were assessed.Constructs expressing miR-584-5p and RUNX2 were established to determine PSC osteogenic differentiation.RESULTS Hypoxic stimulation induced PSC osteogenic differentiation and significantly increased calcified nodules,intracellular calcium ion levels,and alkaline phosphatase(ALP)activity in PSCs.Osteogenic differentiation-related factors such as RUNX2,bone morphogenetic protein 2,hypoxia-inducible factor 1-alpha,and ALP were upregulated;in contrast,miR-584-5p was downregulated in these cells.Furthermore,upregulation of miR-584-5p significantly inhibited RUNX2 expression and hypoxia-induced PSC osteogenic differentiation.RUNX2 was the target gene of miR-584-5p,antagonizing miR-584-5p inhibition in hypoxia-induced PSC osteogenic differentiation.CONCLUSION Our study showed that the interaction of miR-584-5p and RUNX2 could mediate PSC osteogenic differentiation induced by hypoxia.展开更多
Among APETALA2 (AP2)-type plant specific transcription factor family, WRINKLED1 (WRI1), has appeared to be a master gene transcriptionally regulating a set of carbon metabolism- and fatty acid synthesis (FAS)-related ...Among APETALA2 (AP2)-type plant specific transcription factor family, WRINKLED1 (WRI1), has appeared to be a master gene transcriptionally regulating a set of carbon metabolism- and fatty acid synthesis (FAS)-related genes responsible for seed specific triacylglycerols (TAGs) storage in oil plants. B3 type transcription factors, such as ABI3 and FUS3, are known to be involved in seed development, such as seed storage protein synthesis and maturation. Based on the recent whole genome sequence data of castor bean (Ricinus communis L.), putative WRI1 homologs (RcWRI1, RcWRI2) specifically expressed in castor bean seed have been identified by comparing organ specific expression profiles among seed development-related transcription factors, seed storage specific genes (Ricin, RcOleosin) and a set of FAS genes including genes for sucrose synthase (RcSUS2), biotin carboxyl carrier protein (a subunit of acetyl-CoA carboxylase, RcBCCP2) and ketoacyl-acyl carrier protein synthase (RcKAS1). Immunoreactive signals with WRI1, FUS3 and ABI5-related polypeptides were also detected in seed specifically, consistent with the expression profiles of seed development-related genes. The WRI1 binding consensus sites, [CnTnG](n)(7)[CG], designated as the AW-box, were found at the promoter region of RcBCCP2 and RcKAS1. Thus, RcWRI1 possibly play a pivotal role in seed specific TAGs storage during seed development by directly activating FAS -related genes.展开更多
Members of the ERF Family of Transcription Factors play an important role in plant development and gene expression that regulates responses to biotic and abiotic stress.This work identified 36 ERF family genes in Coff...Members of the ERF Family of Transcription Factors play an important role in plant development and gene expression that regulates responses to biotic and abiotic stress.This work identified 36 ERF family genes in Coffea arabica within the AP2/ERF full domain,using the EST-based genomic resource of the Brazilian Coffee Genome Project.The ERF family genes were classified into nine of the ten existing groups through phylogenetic analysis of the deduced amino acid sequences and comparison with the sequences of the ERF family genes in Arabidopsis.In addition to the AP2 domain,other conserved domains were identified,typical of members of each group.The in silico analysis and expression profiling showed high levels of expression for libraries derived from tissues of fruits,leaves and flowers as well as for libraries subjected to water stress.These results suggest the participation of the ERF family genes of C.arabica in distinct biological functions,such as control of development,maturation,and responses to water stress.The results of this work imply in the selection of promising genes for further functional characterizations that will provide a better understanding of the complex regulatory networks related to plant development and responses to stress,opening up opportunities for coffee breeding programs.展开更多
AP2/ERE-type transcription factors,as a type of plant-specific transcription factors,play a key role in plant biotic and abiotic stress.Meanwhile,they have been studied in many plants,but rarely in tomatoes.In this st...AP2/ERE-type transcription factors,as a type of plant-specific transcription factors,play a key role in plant biotic and abiotic stress.Meanwhile,they have been studied in many plants,but rarely in tomatoes.In this study,we performed a genome-wide analysis of the SlAP2/ERF gene family of tomato,and finally identified 29 SlAP2/ERF genes and divided them into different subfamilies.At the same time,its basic physical and chemical properties were analyzed.We also constructed phylogenetic trees with 30 Arabidopsis AP2/ERF proteins and 28 potatoes AP2/ERF proteins to ensure conservative homology between them.In addition,we mapped 29 SlAP2/ERF transcription factors on 10 different chromosomes,and identified 43 responsive plant hormones,responsive light signals,tissue-specific expression and stress response elements from 2000bp upstream of the promoter region,and we analyzed conserved motifs and gene structures of SlAP2/ERF.The tertiary structure of SlAP2/ERF protein was constructed by homology modeling,and the protein-protein interaction network was constructed based on Arabidopsis Thaliana.Finally,the expression pattern of tomato in different tissues was studied by using gene expression database,and the expression level of tomato under abiotic stress was detected by q-RT-PCR.These results provide comprehensive information for further study of the function of the SlAP2/ERF gene family.展开更多
RAV(Related to ABI3/VP1)transcription factors are unique members of the AP2-ERF superfamily with AP2 and B3 domains and play important roles in the regulation of seed germination,plant growth,and stress response.In th...RAV(Related to ABI3/VP1)transcription factors are unique members of the AP2-ERF superfamily with AP2 and B3 domains and play important roles in the regulation of seed germination,plant growth,and stress response.In the study,7 RAV genes,named HvRAVs,were identified in barley based on the available genome sequences.While five of the seven HvRAVs were located on chromosome 3,HvRAV5 and HvRAV7 were located on chromosome 1 and 4,respectively.Six of the predicted HvRAVs were intron-less,except HvRAV2,which had one intron.HvRAV proteins have shown basic,instable,and hydrophilic properties.The AP2 domain specific RAYD and WLG motifs were detected in all HvRAV proteins.Besides,B3 repression domain,R/KLFGV,is also found in the C-terminal of HvRAVs.HvRAVs were found to have stress-related cis-acting elements,including MYB,MYC,and W-BOX.HvRAV2 was predicted to have no GARE motifs,TATCCCA or TAACAA(G/A),and LTREs.Under drought conditions,the expression level of HvRAVs did not significantly change in a drought-sensitive barley genotype,whereas HvRAV5 and HvRAV7 were dramatically down-regulated in a drought-tolerant genotype.Expression of HvRAV5 was also inhibited by salinity.HvRAV7 was strongly induced by plant pathogen attack.Only HvRAV6 was induced by exogenous gibberellin application and during the germination process.Interestingly,HvRAV6 transcript was detected higher than other HvRAVs in all stress and control conditions as well as during germination.In silico analyses have shown that HvRAVs play a role in response to different abiotic and biotic stresses as well as in plant development.However,the extensive biological roles of HvRAV genes in plant development and in response to abiotic and biotic stresses need further investigation.展开更多
Ethylene response factors （ERFs） play important roles in response to plant biotic and abiotic stresses. In this study, a gene encoding a putative AP2/ERF domain-containing protein was isolated by screening a SSH cDN...Ethylene response factors （ERFs） play important roles in response to plant biotic and abiotic stresses. In this study, a gene encoding a putative AP2/ERF domain-containing protein was isolated by screening a SSH cDNA library from rice and designated as Oryza sativa AP2/ERF-like protein （OsAP2LP） gene. OsAP2LP is 1491 bp in length, interrupted by seven introns, and encodes a putative protein of 348 amino acids. Temporal and spatial expression analysis showed that the OsAP2LP gene was preferentially expressed in roots, panicles, mature embryos and seeds in rice. Real-time quantitative PCR analysis indicated that the expression levels of the OsAP2LP gene were increased under the treatments of drought and gibberellin but decreased under the treatments of low temperature, salt, abscisic acid （ABA） and zeatin. Taken together, these results suggest that OsAP2LP might be involved in stress responses, and probably plays roles as a transcription regulator when plants response to cold, salt and drought stresses through ABA and gibberellin pathways.展开更多
基金Supported by Sailing Program of Naval Medical University,Program of Shanghai Hongkou District Health Commission,No.2202-27Special Funds for Activating Scientific Research of Shanghai Fourth People’s Hospital,No.sykyqd05801.
文摘BACKGROUND The hypoxic environment during bone healing is important in regulating the differentiation of periosteal stem cells(PSCs)into osteoblasts or chondrocytes;however,the underlying mechanisms remain unclear.AIM To determine the effect of hypoxia on PSCs,and the expression of microRNA-584-5p(miR-584-5p)and RUNX family transcription factor 2(RUNX2)in PSCs was modulated to explore the impact of the miR-584-5p/RUNX2 axis on hypoxiainduced osteogenic differentiation of PSCs.METHODS In this study,we isolated primary mouse PSCs and stimulated them with hypoxia,and the characteristics and functional genes related to PSC osteogenic differentiation were assessed.Constructs expressing miR-584-5p and RUNX2 were established to determine PSC osteogenic differentiation.RESULTS Hypoxic stimulation induced PSC osteogenic differentiation and significantly increased calcified nodules,intracellular calcium ion levels,and alkaline phosphatase(ALP)activity in PSCs.Osteogenic differentiation-related factors such as RUNX2,bone morphogenetic protein 2,hypoxia-inducible factor 1-alpha,and ALP were upregulated;in contrast,miR-584-5p was downregulated in these cells.Furthermore,upregulation of miR-584-5p significantly inhibited RUNX2 expression and hypoxia-induced PSC osteogenic differentiation.RUNX2 was the target gene of miR-584-5p,antagonizing miR-584-5p inhibition in hypoxia-induced PSC osteogenic differentiation.CONCLUSION Our study showed that the interaction of miR-584-5p and RUNX2 could mediate PSC osteogenic differentiation induced by hypoxia.
文摘Among APETALA2 (AP2)-type plant specific transcription factor family, WRINKLED1 (WRI1), has appeared to be a master gene transcriptionally regulating a set of carbon metabolism- and fatty acid synthesis (FAS)-related genes responsible for seed specific triacylglycerols (TAGs) storage in oil plants. B3 type transcription factors, such as ABI3 and FUS3, are known to be involved in seed development, such as seed storage protein synthesis and maturation. Based on the recent whole genome sequence data of castor bean (Ricinus communis L.), putative WRI1 homologs (RcWRI1, RcWRI2) specifically expressed in castor bean seed have been identified by comparing organ specific expression profiles among seed development-related transcription factors, seed storage specific genes (Ricin, RcOleosin) and a set of FAS genes including genes for sucrose synthase (RcSUS2), biotin carboxyl carrier protein (a subunit of acetyl-CoA carboxylase, RcBCCP2) and ketoacyl-acyl carrier protein synthase (RcKAS1). Immunoreactive signals with WRI1, FUS3 and ABI5-related polypeptides were also detected in seed specifically, consistent with the expression profiles of seed development-related genes. The WRI1 binding consensus sites, [CnTnG](n)(7)[CG], designated as the AW-box, were found at the promoter region of RcBCCP2 and RcKAS1. Thus, RcWRI1 possibly play a pivotal role in seed specific TAGs storage during seed development by directly activating FAS -related genes.
文摘Members of the ERF Family of Transcription Factors play an important role in plant development and gene expression that regulates responses to biotic and abiotic stress.This work identified 36 ERF family genes in Coffea arabica within the AP2/ERF full domain,using the EST-based genomic resource of the Brazilian Coffee Genome Project.The ERF family genes were classified into nine of the ten existing groups through phylogenetic analysis of the deduced amino acid sequences and comparison with the sequences of the ERF family genes in Arabidopsis.In addition to the AP2 domain,other conserved domains were identified,typical of members of each group.The in silico analysis and expression profiling showed high levels of expression for libraries derived from tissues of fruits,leaves and flowers as well as for libraries subjected to water stress.These results suggest the participation of the ERF family genes of C.arabica in distinct biological functions,such as control of development,maturation,and responses to water stress.The results of this work imply in the selection of promising genes for further functional characterizations that will provide a better understanding of the complex regulatory networks related to plant development and responses to stress,opening up opportunities for coffee breeding programs.
基金the National Natural Science Foundation of China(31560663,32060401).
文摘AP2/ERE-type transcription factors,as a type of plant-specific transcription factors,play a key role in plant biotic and abiotic stress.Meanwhile,they have been studied in many plants,but rarely in tomatoes.In this study,we performed a genome-wide analysis of the SlAP2/ERF gene family of tomato,and finally identified 29 SlAP2/ERF genes and divided them into different subfamilies.At the same time,its basic physical and chemical properties were analyzed.We also constructed phylogenetic trees with 30 Arabidopsis AP2/ERF proteins and 28 potatoes AP2/ERF proteins to ensure conservative homology between them.In addition,we mapped 29 SlAP2/ERF transcription factors on 10 different chromosomes,and identified 43 responsive plant hormones,responsive light signals,tissue-specific expression and stress response elements from 2000bp upstream of the promoter region,and we analyzed conserved motifs and gene structures of SlAP2/ERF.The tertiary structure of SlAP2/ERF protein was constructed by homology modeling,and the protein-protein interaction network was constructed based on Arabidopsis Thaliana.Finally,the expression pattern of tomato in different tissues was studied by using gene expression database,and the expression level of tomato under abiotic stress was detected by q-RT-PCR.These results provide comprehensive information for further study of the function of the SlAP2/ERF gene family.
文摘RAV(Related to ABI3/VP1)transcription factors are unique members of the AP2-ERF superfamily with AP2 and B3 domains and play important roles in the regulation of seed germination,plant growth,and stress response.In the study,7 RAV genes,named HvRAVs,were identified in barley based on the available genome sequences.While five of the seven HvRAVs were located on chromosome 3,HvRAV5 and HvRAV7 were located on chromosome 1 and 4,respectively.Six of the predicted HvRAVs were intron-less,except HvRAV2,which had one intron.HvRAV proteins have shown basic,instable,and hydrophilic properties.The AP2 domain specific RAYD and WLG motifs were detected in all HvRAV proteins.Besides,B3 repression domain,R/KLFGV,is also found in the C-terminal of HvRAVs.HvRAVs were found to have stress-related cis-acting elements,including MYB,MYC,and W-BOX.HvRAV2 was predicted to have no GARE motifs,TATCCCA or TAACAA(G/A),and LTREs.Under drought conditions,the expression level of HvRAVs did not significantly change in a drought-sensitive barley genotype,whereas HvRAV5 and HvRAV7 were dramatically down-regulated in a drought-tolerant genotype.Expression of HvRAV5 was also inhibited by salinity.HvRAV7 was strongly induced by plant pathogen attack.Only HvRAV6 was induced by exogenous gibberellin application and during the germination process.Interestingly,HvRAV6 transcript was detected higher than other HvRAVs in all stress and control conditions as well as during germination.In silico analyses have shown that HvRAVs play a role in response to different abiotic and biotic stresses as well as in plant development.However,the extensive biological roles of HvRAV genes in plant development and in response to abiotic and biotic stresses need further investigation.
基金supported by the National Natural Science Foundation of China (Grant Nos.30770132 and 30570103)
文摘Ethylene response factors （ERFs） play important roles in response to plant biotic and abiotic stresses. In this study, a gene encoding a putative AP2/ERF domain-containing protein was isolated by screening a SSH cDNA library from rice and designated as Oryza sativa AP2/ERF-like protein （OsAP2LP） gene. OsAP2LP is 1491 bp in length, interrupted by seven introns, and encodes a putative protein of 348 amino acids. Temporal and spatial expression analysis showed that the OsAP2LP gene was preferentially expressed in roots, panicles, mature embryos and seeds in rice. Real-time quantitative PCR analysis indicated that the expression levels of the OsAP2LP gene were increased under the treatments of drought and gibberellin but decreased under the treatments of low temperature, salt, abscisic acid （ABA） and zeatin. Taken together, these results suggest that OsAP2LP might be involved in stress responses, and probably plays roles as a transcription regulator when plants response to cold, salt and drought stresses through ABA and gibberellin pathways.
