Ward M J, Mok K C, Astling D P, Lew H, Zusman D R
Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3204, USA.
J Bacteriol. 1998 Nov;180(21):5697-703. doi: 10.1128/JB.180.21.5697-5703.1998.
Myxococcus xanthus is a gram-negative bacterium which has a complex life cycle. Autochemotaxis, a process whereby cells release a self-generated signaling molecule, may be the principal mechanism facilitating directed motility in both the vegetative swarming and developmental aggregation stages of this life cycle. The process requires the Frz signal transduction system, including FrzZ, a protein which is composed of two domains, both showing homology to the enteric chemotaxis response regulator CheY. The first domain of FrzZ (FrzZ1), when expressed as bait in the yeast two-hybrid system and screened against a library, was shown to potentially interact with the C-terminal portion of a protein encoding an ATP-binding cassette (AbcA). The activation domain-AbcA fusion protein did not interact with the second domain of FrzZ (FrzZ2) or with two other M. xanthus response regulator-containing proteins presented as bait, suggesting that the FrzZ1-AbcA interaction may be specific. Cloning and sequencing of the upstream region of the abcA gene showed the ATP-binding cassette to be linked to a large hydrophobic, potentially membrane-spanning domain. This domain organization is characteristic of a subgroup of ABC transporters which perform export functions. Cloning and sequencing downstream of abcA indicated that the ABC transporter is at the start of an operon containing three open reading frames. An insertion mutation in the abcA gene resulted in cells displaying the frizzy aggregation phenotype, providing additional evidence that FrzZ and AbcA may be part of the same signal transduction pathway. Cells with mutations in genes downstream of abcA showed no developmental defects. Analysis of the proposed exporter role of AbcA in cell mixing experiments showed that the ABC transporter mutant could be rescued by extracellular complementation. We speculate that the AbcA protein may be involved in the export of a molecule required for the autochemotactic process.
黄色粘球菌是一种革兰氏阴性细菌,具有复杂的生命周期。自身趋化作用是细胞释放自身产生的信号分子的过程,可能是在该生命周期的营养群体运动和发育聚集阶段促进定向运动的主要机制。这个过程需要Frz信号转导系统,包括FrzZ,一种由两个结构域组成的蛋白质,这两个结构域都与肠道趋化反应调节因子CheY具有同源性。当在酵母双杂交系统中作为诱饵表达并针对文库进行筛选时,FrzZ的第一个结构域(FrzZ1)显示可能与编码ATP结合盒(AbcA)的蛋白质的C末端部分相互作用。激活结构域-AbcA融合蛋白不与FrzZ的第二个结构域(FrzZ2)或作为诱饵呈现的其他两种含黄色粘球菌反应调节因子的蛋白质相互作用,这表明FrzZ1-AbcA相互作用可能是特异性的。abcA基因上游区域的克隆和测序表明,ATP结合盒与一个大的疏水、可能跨膜的结构域相连。这种结构域组织是执行输出功能的ABC转运蛋白亚组的特征。abcA下游的克隆和测序表明,ABC转运蛋白位于一个包含三个开放阅读框的操纵子的起始位置。abcA基因中的插入突变导致细胞呈现卷曲聚集表型,这提供了额外的证据表明FrzZ和AbcA可能是同一信号转导途径的一部分。abcA下游基因发生突变的细胞没有显示出发育缺陷。在细胞混合实验中对AbcA拟议的输出作用进行分析表明,ABC转运蛋白突变体可以通过细胞外互补得到拯救。我们推测,AbcA蛋白可能参与自身趋化过程所需分子的输出。