Systematic analysis, by the yeast two-hybrid, of protein interaction between components of the type II secretory machinery of Erwinia chrysanthemi

Abstract

Type II systems allow for the secretion of numerous enzymes and toxins in several Gram-negative pathogens. In Erwinia chrysanthemi, 14 Out proteins are necessary for building the type II apparatus. We performed a systematic two-hybrid analysis to test interactions between the periplasmic regions of the Out proteins. Results obtained using this approach suggested that OutJ (a pseudopilin) was able to interact with (i) OutD, the outer membrane secretin, (ii) OutI, mainly located in the periplasm, and (iii) OutL, an inner membrane protein. Taken together, these results suggest that OutJ is involved in multiple partnerships. Implications of these partnerships in the overall architecture of the type II secretion machinery are discussed.