Chung Y S, Dubnau D
Public Health Research Institute, New York, New York 10016, USA.
Mol Microbiol. 1995 Feb;15(3):543-51. doi: 10.1111/j.1365-2958.1995.tb02267.x.
ComGC is a cell surface-localized protein required for DNA binding during transformation in Bacillus subtilis. It resembles type IV prepilins in its N-terminal domain, particularly in the amino acid sequence surrounding the processing cleavage sites of these proteins. ComC is another protein required for DNA binding, which resembles the processing proteases that cleave type IV prepilins. We show here that ComGC is processed in competent cells and that this processing requires ComC. We also demonstrate that the PilD protein of Neisseria gonorrhoeae, a ComC homologue, can process ComGC in Escherichia coli, and that the ComC protein itself is the only B. subtilis protein needed to accomplish cleavage of ComGC in the latter organism. Based on NaOH-solubility studies, we have shown that in the absence of ComC, but in the presence of all other competence proteins, B. subtilis is incapable of correctly translocating ComGC to the outer face of the cell membrane. Finally, we show that ComGC can be cross-linked to yield a form with higher molecular mass, possibly a dimer, and present evidence suggesting that formation of the higher mass complex takes place in the membrane, prior to translocation. Formation of this complex does not require ComC or any of the comG products, other than ComGC itself.
ComGC是枯草芽孢杆菌转化过程中DNA结合所需的一种细胞表面定位蛋白。它在N端结构域类似于IV型前菌毛蛋白,特别是在这些蛋白加工切割位点周围的氨基酸序列方面。ComC是另一种DNA结合所需的蛋白,它类似于切割IV型前菌毛蛋白的加工蛋白酶。我们在此表明,ComGC在感受态细胞中被加工,且这种加工需要ComC。我们还证明,淋病奈瑟菌的PilD蛋白(一种ComC同源物)可以在大肠杆菌中加工ComGC,并且ComC蛋白本身是枯草芽孢杆菌中完成ComGC切割所需的唯一蛋白。基于氢氧化钠溶解性研究,我们表明,在没有ComC但存在所有其他感受态蛋白的情况下,枯草芽孢杆菌无法将ComGC正确转运到细胞膜外表面。最后,我们表明ComGC可以交联形成分子量更高的形式,可能是二聚体,并提供证据表明更高质量复合物的形成发生在转运之前的膜中。这种复合物的形成不需要ComC或任何comG产物,除了ComGC本身。