Demonstration of interactions among Myxococcus xanthus Dif chemotaxis-like proteins by the yeast two-hybrid system.
Demonstration of interactions among Myxococcus xanthus Dif chemotaxis-like proteins by the yeast two-hybrid system.
复制标题
通过酵母双杂交系统证明黄色粘球菌 Dif 趋化样蛋白之间的相互作用。
DOI：
10.1007/s00203-005-0767-8
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Li,Zhuo
中科院分区：
文献类型：
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作者：
Yang,Zhaomin;Li,Zhuo
黄色粘球菌编码几种细菌趋化性同源物，对纤维胞外多糖(EPS)的产生、社会滑动运动和子实体发育至关重要。在一级序列中，DIFA与甲基接受趋化蛋白同源，DIFC与咀嚼同源，DIFD与Chey同源，DifA与CHEA同源，DifG与Check同源。在本研究中，利用酵母双杂交(Y2H)系统研究了类Dif趋化蛋白之间的相互作用。DIFC被发现同时与DIFA和DIFE相互作用。利用Y2H或“三杂”体系，证明了DIFC能够调节DIFA、DIFC和DIFE三元蛋白质复合体的形成。根据序列分析，DIFE的保守结构域可能反映了Chea-type激酶的功能保守性，因为它的P2结构域与Difd相互作用，P5与DIFC相互作用，而P3结构域似乎是二聚化的。类似地，difa的C-末端区域似乎也是二聚体。此外，还发现DifG与Difd相互作用，这与DifG是Difd-磷酸的一种磷酸酶的假说是一致的。这些发现支持这样的模型，即Dif蛋白构成了一个独特的趋化性信号转导途径，在调节EPS产生INM方面具有中心功能。黄花草。
TheMyxococcus xanthus diflocus encodes several bacterial chemotaxis homologues that are crucial for fibril exopolysaccharide (EPS) production, social gliding motility, and fruiting body development. In primary sequence, DifA is homologous to methyl-accepting chemotaxis protein, DifC to CheW, DifD to CheY, DifE to CheA, and DifG to CheC. In this study, the interactions among the Dif chemotaxis-like proteins were investigated using the yeast two-hybrid (Y2H) system. DifC was found to interact with both DifA and DifE. Using a modified Y2H or a “three-hybrid” system, it was demonstrated that DifC is capable of mediating the formation of DifA, DifC, and DifE ternary protein complexes. The conserved domains of DifE, based on sequence analysis, likely reflect functional conservations of CheA-type kinases, because its P2 domain interacts with DifD, P5 with DifC, and the P3 domain appears to dimerize. Similarly, C-terminal regions of DifA appear to dimerize as well. In addition, DifG was found to interact with DifD, which is consistent with the hypothesis that DifG is a phosphatase of DifD-phosphate. These findings support the models in which Dif proteins constitute a unique chemotaxis-like signal transduction pathway with central functions in regulating EPS production inM. xanthus.