A non-polar mutation in bfpE will be constructed, and the effect of this lesion on pilus production and bacterial adherence will be examined. A detailed topological analysis of BfpA will be carried out using phoA/lacZ reporter systems. The effects of mutations in the hydrophilic segments of BfpE will be determined to identify critical residues for pilus function. Three different methods: selection of suppressor of BfpE, cross linking, and use of the two-hybrid system, are proposed to study the interaction of BfpE with other proteins.
The BfpE protein of enteropathogenic E. coli (EPEC) is a representative of a family of cytoplasmic membrane proteins that participate in the biogenesis of type IV pili. It is hypothesized that BfpE is directly required for pilus formation and that it is a key component of a protein complex encoded by the bfp gene cluster. Experimental strategies are proposed to gain a better understanding of the structure and function of BfpE.
<p>
The results of the proposed research should advance our knowledge about the mechanisms involved in the biogenesis of EPEC pili.