simple sequence repeats and mucoid conversion biased muca mutagenesis in mismatch repair-deficient pseudomonas aeruginosa简单序列重复和黏液状的转换偏见muca诱变在失配repair-deficient铜绿假单胞菌.pdfVIP

Simple Sequence Repeats and Mucoid Conversion: Biased mucA Mutagenesis in Mismatch Repair-Deficient Pseudomonas aeruginosa Alejandro J. Moyano, Andrea M. Smania* ´ ´ ´ ´ ´ ´ Centro de Investigaciones en Quımica Biologica de Cordoba (CIQUIBIC), CONICET, Departamento de Quımica Biologica, Facultad de Ciencias Quımicas, Universidad ´ ´ Nacional de Cordoba, Cordoba, Argentina Abstract In Pseudomonas aeruginosa, conversion to the mucoid phenotype marks the onset of an irreversible state of the infection in Cystic Fibrosis (CF) patients. The main pathway for mucoid conversion is mutagenesis of the mucA gene, frequently due to 21 bp deletions in a simple sequence repeat (SSR) of 5 Gs (G5-SSR426). We have recently observed that this mucA mutation is particularly accentuated in Mismatch Repair System (MRS)-deficient cells grown in vitro. Interestingly, previous reports have shown a high prevalence of hypermutable MRS-deficient strains occurring naturally in CF chronic lung infections. Here, we used mucA as a forward mutation model to systematically evaluate the role of G5-SSR426 in conversion to mucoidy in a MRS-deficient background, with this being the first analysis combining SSR-dependent localized hypermutability and the acquisition of a particular virulence/persistence trait in P. aeruginosa. In this study, mucA alleles were engineered with different contents of G:C SSRs, and tested for their effect on the mucoid conversion frequency and mucA mutational spectra in a mutS-deficient strain of P. aeruginosa. Importantly, deletion of G5-SSR426 severely reduced the emergence frequency of mucoid variants, with no preferential site of mutagenesis within mucA. Moreover, although mutagenesis in muc