1887

Abstract

A number of single-nucleotide polymorphisms (SNPs) have been identified in the genome of BCG Pasteur compared with the sequenced strain 2122/97. The functional consequences of many of these mutations remain to be described; however, mutations in genes encoding regulators may be particularly relevant to global phenotypic changes such as loss of virulence, since alteration of a regulator's function will affect the expression of a wide range of genes. One such SNP falls in , encoding a member of the AfsR/DnrI/SARP class of global transcriptional regulators, that replaces a highly conserved glutamic acid residue at position 159 (E159G) with glycine in a tetratricopeptide repeat (TPR) located in the bacterial transcriptional activation (BTA) domain of BCG3145. TPR domains are associated with protein–protein interactions, and a conserved core (helices T1–T7) of the BTA domain seems to be required for proper function of SARP-family proteins. Structural modelling predicted that the E159G mutation perturbs the third -helix of the BTA domain and could therefore have functional consequences. The E159G SNP was found to be present in all BCG strains, but absent from virulent and strains. By overexpressing BCG3145 and Rv3124 in BCG and H37Rv and monitoring transcriptome changes using microarrays, we determined that BCG3145/Rv3124 acts as a positive transcriptional regulator of the molybdopterin biosynthesis locus, and we suggest that be renamed . The SNP in was found to have a subtle effect on the activity of MoaR1, suggesting that this mutation is not a key event in the attenuation of BCG.

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2010-07-01
2019-10-19
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vol. , part 7, pp. 2112 - 2123

[ PDF, 87 kb], including: Primers used for qRT-PCR Plasmids and primers used for the generation of mutated versions of qRT-PCR fold-changes of Rv3124 and the locus in BCG/pB1 versus BCG/pB3 Southern blot analysis of BCG mutant CLUSTAL alignment of members of the AfsR/DnrI/SARP family



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