Cantwell Brian J, Draheim Roger R, Weart Richard B, Nguyen Cameran, Stewart Richard C, Manson Michael D
Department of Biology, Texas A&M University, College Station, Texas 77843, USA.
J Bacteriol. 2003 Apr;185(7):2354-61. doi: 10.1128/JB.185.7.2354-2361.2003.
We have investigated the conditions required for polar localization of the CheZ phosphatase by using a CheZ-green fluorescent protein fusion protein that, when expressed from a single gene in the chromosome, restored chemotaxis to a DeltacheZ strain. Localization was observed in wild-type, DeltacheZ, DeltacheYZ, and DeltacheRB cells but not in cells with cheA, cheW, or all chemoreceptor genes except aer deleted. Cells making only CheA-short (CheA(S)) or CheA lacking the P2 domain also retained normal localization, whereas cells producing only CheA-long or CheA missing the P1 and P2 domains did not. We conclude that CheZ localization requires the truncated C-terminal portion of the P1 domain present in CheA(S). Missense mutations targeting residues 83 through 120 of CheZ also abolished localization. Two of these mutations do not disrupt chemotaxis, indicating that they specifically prevent interaction with CheA(S) while leaving other activities of CheZ intact.
我们通过使用CheZ-绿色荧光蛋白融合蛋白研究了CheZ磷酸酶极性定位所需的条件,该融合蛋白从染色体上的单个基因表达时,可恢复趋化性至ΔcheZ菌株。在野生型、ΔcheZ、ΔcheYZ和ΔcheRB细胞中观察到定位,但在除aer外的cheA、cheW或所有化学感受器基因缺失的细胞中未观察到。仅产生CheA-short(CheA(S))或缺失P2结构域的CheA的细胞也保留正常定位,而仅产生CheA-long或缺失P1和P2结构域的CheA的细胞则没有。我们得出结论,CheZ定位需要CheA(S)中存在的P1结构域的截短C末端部分。针对CheZ的83至120位残基的错义突变也消除了定位。其中两个突变不会破坏趋化性,表明它们特异性地阻止与CheA(S)的相互作用,同时保持CheZ的其他活性完整。