Submitted to: Molecular Microbiology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 6/1/1998
Publication Date: N/A
Citation: Chung, Y.S., Breidt, F., Dubnau, D. 1998. Cell surface localization and processing of the ComG proteins, required for DNA binding during transformation of bacillus subtilis. Mol. Microbiol. 29:905-913. Interpretive Summary: This paper describes a characterization of structures in the cell membrane of a common soil bacterium (Bacillus subtilis) that functions to allow the cell to take up DNA from the environment. While it has been known for many years that certain bacteria are capable of the uptake of DNA, the mechanism by which this process occurs is largely unexplored. This paper describes an investigation of the molecular mechanism by which DNA uptake occurs. Commonly used tools of immunology and molecular biology were used to study the relationship and function of the proteins involved and the genes encoding these proteins. The results described in the paper show how some of the proteins interact and how these proteins are positioned in the bacterial cell membrane. The significance of the work is primarily to advance our basic understanding of this important process. Understanding this mechanism may lead to new insights in evolutionary theory relating to bacteria. The exchange of genes by pathogenic bacteria, using an uptake mechanism similar to the one investigated in this work, may contribute to the occurrence of virulent new bacterial strains that are difficult to kill with antibiotics.
Technical Abstract: The comG operon of Bacillus subtilis encodes seven proteins essential for the binding of transforming DNA to the competent cell surface. We have explored the processing of the ComG proteins and the cellular localization of six of them. All of the proteins were found to be membrane associated. The four proteins with N-terminal sequence motifs typical of type 4 prepilins (ComGC, GD, GE and GG) are processed by a pathway that requires the product of comC, also an essential competence gene. The unprocessed forms of ComGC and GD behave like integral membrane proteins. Pre-ComGG differs from pre-ComGC and pre-ComGD, in that it is accessible to proteolysis only from the cytoplasmic face of the membrane and at least a portion of it behaves like a peripheral membrane protein. The mature forms of these proteins are translocated to the outer face of the membrane and are liberated when peptidoglycan is hydrolysed by lysozyme or mutanolysin. ComGG exists in part as a disulphide-cross-linked homodimer in vivo. ComGC was found to possess an intramolecular disulphide bond. The previously identified homodimer form of this protein is not stabilized by disulphide bond formation. ComGF behaves as an integral membrane protein, while ComGA, a putative ATPase, is located on the inner face of the membrane as a peripheral membrane protein. Possible roles of the ComG proteins in DNA binding to the competent cell surface are discussed in the light of these and other results.