Environmental control of the alcohol dehydrogenase(Adh)and other stress response genes in plants is in part brought about by transcriptional regulation involving the G-box cis-acting DNA element and bZIP G-box Binding...Environmental control of the alcohol dehydrogenase(Adh)and other stress response genes in plants is in part brought about by transcriptional regulation involving the G-box cis-acting DNA element and bZIP G-box Binding Factors(GBFs).The mechanisms of GBF regulation and requirements for additional factors in this control process are not well understood.In an effort to identify potential GBF binding and control partners,maize GBF1 was used as bait in a yeast two-hybrid screen of an A.thaliana cDNA library.GBF Interacting Protein 1(GIP1)arose from the screen as a 496 amino acid protein with a predicted molecular weight of 53,748 kDa that strongly interacts with GBFs.Northern analysis of A.thaliana tissue suggests a 1.8-1.9 kb GIP1 transcript,predominantly in roots.Immunolocalization studies indicate that GIP1 protein is mainly localized to the nucleus.In vitro electrophoretic mobility shift assays using an Adh G-box DNA probe and recombinant A.thaliana GBF3 or maize GBF1,showed that the presence of GIP1 resulted in a tenfold increase in GBF DNA binding activity without altering the migration,suggesting a transient association between GIP1 and GBF.Addition of GIP1 to intentionally aggregated GBF converted GBF to lower molecular weight macromolecular complexes and GIP1 also refolded denatured rhodanese in the absence of ATP.These data suggest GIP1 functions to enhance GBF DNA binding activity by acting as a potent nuclear chaperone or crowbar,and potentially regulates the multimeric state of GBFs,thereby contributing to bZIP-mediated gene regulation.展开更多
A new hypocrellin B （HB） derivative bearing a bispyrrolecarboxamide-containing side chain was synthesized, which presents improved absorptivity in phototherapeufic window than HB and affinity feature towards dsDNA.
Objective:Salvia miltiorrhiza is widely used in traditional Chinese medicine for treating cardiovascular and cerebrovascular diseases,with tanshinones being its major active components.This study aims to systematicall...Objective:Salvia miltiorrhiza is widely used in traditional Chinese medicine for treating cardiovascular and cerebrovascular diseases,with tanshinones being its major active components.This study aims to systematically elucidate the core transcriptional circuitry controlling tanshinone production,thereby establishing a mechanistic framework to optimize phytochemical yield and advance sustainable cultivation strategies for this pharmaceutically vital species.Methods:Transcriptome profiling revealed that the transcription factor SmWRKY69 is specifically expressed in the root periderm of S.miltiorrhiza.DNA affinity purification sequencing(DAPseq)was used to identify its potential target genes,and cis-element analysis predicted W-box motifs in the promoters of SmCPS1 and SmKSL1.Yeast one-hybrid(Y1H)assays were employed to validate its regulatory interactions with candidate gene promoters.Results:SmWRKY69 was found to directly bind to the promoters of SmCPS1 and SmKSL1,key genes in the tanshinone biosynthetic pathway,through W-box elements,indicating its role as a transcriptional regulator.Conclusion:SmWRKY69 regulates tanshinone biosynthesis by directly targeting SmCPS1 and SmKSL1,providing a valuable genetic target for metabolic engineering to enhance the therapeutic quality of S.miltiorrhiza.展开更多
The DNA binding and cleavage activity of a synthetic polyamide containing dipeptide Ser-His has been investigated by spectroscopic techniques,such as electronic absorbance and circular dichroism(CD)spectropolarimetry,...The DNA binding and cleavage activity of a synthetic polyamide containing dipeptide Ser-His has been investigated by spectroscopic techniques,such as electronic absorbance and circular dichroism(CD)spectropolarimetry,as well as gel electrophoresis.The results show that the molecule has a strong interaction with DNA and can improve DNA-cleavage ability about 100 fold as compared with single dipeptide Ser-His.展开更多
基金This research was supported by the U S Department of Agriculture Grants 00-35304-96Ol and 98-35301-6083.
文摘Environmental control of the alcohol dehydrogenase(Adh)and other stress response genes in plants is in part brought about by transcriptional regulation involving the G-box cis-acting DNA element and bZIP G-box Binding Factors(GBFs).The mechanisms of GBF regulation and requirements for additional factors in this control process are not well understood.In an effort to identify potential GBF binding and control partners,maize GBF1 was used as bait in a yeast two-hybrid screen of an A.thaliana cDNA library.GBF Interacting Protein 1(GIP1)arose from the screen as a 496 amino acid protein with a predicted molecular weight of 53,748 kDa that strongly interacts with GBFs.Northern analysis of A.thaliana tissue suggests a 1.8-1.9 kb GIP1 transcript,predominantly in roots.Immunolocalization studies indicate that GIP1 protein is mainly localized to the nucleus.In vitro electrophoretic mobility shift assays using an Adh G-box DNA probe and recombinant A.thaliana GBF3 or maize GBF1,showed that the presence of GIP1 resulted in a tenfold increase in GBF DNA binding activity without altering the migration,suggesting a transient association between GIP1 and GBF.Addition of GIP1 to intentionally aggregated GBF converted GBF to lower molecular weight macromolecular complexes and GIP1 also refolded denatured rhodanese in the absence of ATP.These data suggest GIP1 functions to enhance GBF DNA binding activity by acting as a potent nuclear chaperone or crowbar,and potentially regulates the multimeric state of GBFs,thereby contributing to bZIP-mediated gene regulation.
文摘A new hypocrellin B （HB） derivative bearing a bispyrrolecarboxamide-containing side chain was synthesized, which presents improved absorptivity in phototherapeufic window than HB and affinity feature towards dsDNA.
文摘Objective:Salvia miltiorrhiza is widely used in traditional Chinese medicine for treating cardiovascular and cerebrovascular diseases,with tanshinones being its major active components.This study aims to systematically elucidate the core transcriptional circuitry controlling tanshinone production,thereby establishing a mechanistic framework to optimize phytochemical yield and advance sustainable cultivation strategies for this pharmaceutically vital species.Methods:Transcriptome profiling revealed that the transcription factor SmWRKY69 is specifically expressed in the root periderm of S.miltiorrhiza.DNA affinity purification sequencing(DAPseq)was used to identify its potential target genes,and cis-element analysis predicted W-box motifs in the promoters of SmCPS1 and SmKSL1.Yeast one-hybrid(Y1H)assays were employed to validate its regulatory interactions with candidate gene promoters.Results:SmWRKY69 was found to directly bind to the promoters of SmCPS1 and SmKSL1,key genes in the tanshinone biosynthetic pathway,through W-box elements,indicating its role as a transcriptional regulator.Conclusion:SmWRKY69 regulates tanshinone biosynthesis by directly targeting SmCPS1 and SmKSL1,providing a valuable genetic target for metabolic engineering to enhance the therapeutic quality of S.miltiorrhiza.
基金Supported by the National Natural Science Foundation of China(Grant No.20572061)
文摘The DNA binding and cleavage activity of a synthetic polyamide containing dipeptide Ser-His has been investigated by spectroscopic techniques,such as electronic absorbance and circular dichroism(CD)spectropolarimetry,as well as gel electrophoresis.The results show that the molecule has a strong interaction with DNA and can improve DNA-cleavage ability about 100 fold as compared with single dipeptide Ser-His.
