An MYB transcription factor (AtMYB305) of Arabidopsis was structurally characterized. The biochemical activity of the protein was tested and its in vivo function was preliminarily analyzed. AtMYB305 contains two imper...An MYB transcription factor (AtMYB305) of Arabidopsis was structurally characterized. The biochemical activity of the protein was tested and its in vivo function was preliminarily analyzed. AtMYB305 contains two imperfect repeats (R2, R3) in its DNA binding domain. Gel mobility shift assay demonstrated that GST-AtMYB305 fusion pro-tein bound specifically to the DNA fragment that included a consensus MYB recognition sequence (TAACTG). Overex-pression of AtMYB305 in the fission yeast caused the forma-tion of elongated cells with one condensed nucleus. Semi-quantitative RT-PCR analysis revealed that AtMYB305 was expressed specifically in the roots of Arabidopsis.展开更多
Our previous study demonstrated that WLIMla has dual roles in fiber elongation and secondary cell wall synthesis in upland cotton, and the protein acts either as an actin-binding protein or as a transcription factor. ...Our previous study demonstrated that WLIMla has dual roles in fiber elongation and secondary cell wall synthesis in upland cotton, and the protein acts either as an actin-binding protein or as a transcription factor. Because WLIMla consists of two different LIM domains, it is possible that these elements contribute differentially to the dual functions of the protein. In this study, we dissected the two LIM domains and characterized their biochemical functions. By using red fluorescent protein （RFP） fusion, co-sedimentation, and DNA binding methods, we found that the two domains of WLIM 1 a, domain 1 （D 1） and domain2 （D2）, possessed different biochemical properties. While D1 contributed primarily to the actin filament-bundling activity of WLIMla, D2 contributed to the DNA-binding activity of the protein; both D1 and D2 relied on a linker sequence for their ac- tivities. In addition, we found that WLIMla and its two LIM domains form dimers in vitro. These results may lead to a better understanding of the molecular mechanisms of dual functions of WLIMla during cotton fiber development.展开更多
文摘An MYB transcription factor (AtMYB305) of Arabidopsis was structurally characterized. The biochemical activity of the protein was tested and its in vivo function was preliminarily analyzed. AtMYB305 contains two imperfect repeats (R2, R3) in its DNA binding domain. Gel mobility shift assay demonstrated that GST-AtMYB305 fusion pro-tein bound specifically to the DNA fragment that included a consensus MYB recognition sequence (TAACTG). Overex-pression of AtMYB305 in the fission yeast caused the forma-tion of elongated cells with one condensed nucleus. Semi-quantitative RT-PCR analysis revealed that AtMYB305 was expressed specifically in the roots of Arabidopsis.
基金the National Basic Research Priorities Program (U1303281)the China Postdoctoral Science Foundation
文摘Our previous study demonstrated that WLIMla has dual roles in fiber elongation and secondary cell wall synthesis in upland cotton, and the protein acts either as an actin-binding protein or as a transcription factor. Because WLIMla consists of two different LIM domains, it is possible that these elements contribute differentially to the dual functions of the protein. In this study, we dissected the two LIM domains and characterized their biochemical functions. By using red fluorescent protein （RFP） fusion, co-sedimentation, and DNA binding methods, we found that the two domains of WLIM 1 a, domain 1 （D 1） and domain2 （D2）, possessed different biochemical properties. While D1 contributed primarily to the actin filament-bundling activity of WLIMla, D2 contributed to the DNA-binding activity of the protein; both D1 and D2 relied on a linker sequence for their ac- tivities. In addition, we found that WLIMla and its two LIM domains form dimers in vitro. These results may lead to a better understanding of the molecular mechanisms of dual functions of WLIMla during cotton fiber development.
