1887

Abstract

A surrogate expression system, based on fusions to the bacterial reporter gene, was used to identify genes that encode exported proteins and the promoter regions required for their expression in the heterologous host . To assess these results in the context of the complete genome sequence, the corresponding genes were identified and computational algorithms were employed to identify signal peptide (SP), transmembrane domain and membrane lipoprotein attachment motifs. This information was used to predict the subset of genes that encode exported proteins. Of the 34 genes identified by the method, 22 were classified to encode potential soluble secreted proteins. Among these, 14 genes may encode novel secreted proteins. Six of the remaining 12 genes were predicted to encode membrane lipoproteins and an additional six to encode integral membrane proteins. Published observations of proteins proven to be secreted into culture filtrates were reviewed to further characterize the mycobacterial SP motif. It was concluded that mycobacterial SPs are comparable in size to Gram-positive SPs, but certain features are different. In particular, arginine was the predominant N-terminally positively charged amino acid in contrast to lysine in the Gram-positives. The hydrophobic transmembrane segment of the SP was dominated by alanine, in contrast to leucine. At the C-terminal end of the SPs, the (−3, −1) rule (AXA motif) holds, with alanine as the dominant amino acid in both positions, being most dominant in the (−1) position. A high proportion of mature sequences start with aspartic acid in the (+1) position and proline in the (+2) position – the DP motif. The authors propose that the DP sequence serves as a sorting signal, following translocation and cleavage by signal peptidase I. Alternatively, the DP motif may be part of the recognition site for the signal peptidase.

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2000-07-01
2019-09-17
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