3). Thus, ydbK is needed for superoxide resistance upon growth
on minimal media but ompN is not. In a second attempt to investigate the ompN function, we then hypothesized that OmpN may play a role in MDR because ompN overexpression was initially found for the MDR mutant NorE5. To assess its function, the ompN gene was cloned into a pUC19 derivative multicopy vector and transformed into PS5 (P-O12). The susceptibility profile of strains PS5, P-O12, and P-9817 (PS5 carrying the vector alone) was assayed for several unrelated antibiotics (norfloxacin, ciprofloxacin, chloramphenicol, tetracycline, erythromycin, trimethoprim, and ceftriaxone). Results showed no significant difference between the strains tested (data not shown). Moreover, the ompN and ydbK mutants (M6131, M6135, and M6133, M6137, respectively) as well as the parental learn more strains (GC4468 and M5950) were similarly tested in MH or M9 agar plates Everolimus despite no significant difference being detected (data not shown). Altogether, these results show no role for this two-gene operon in conferring the MDR phenotype as tested here. In summary, this study has shown that ydbK and ompN are coexpressed in the same mRNA transcript and coordinately activated by SoxS. This activation is exerted on the promoter upstream
of the ydbK gene and presumably results from an indirect effect. Nonetheless, the ydbK gene but not ompN showed a function related to superoxide resistance (only when growing on minimal media), whereas neither function was needed for antimicrobial resistance. Thus, further studies are required to better characterize the ompN function. We wish to thank B. Demple for kindly providing the strains GC4468 and JTG936 and M. M. Tavío for providing the PS5 and NorE5 strains. This study has been supported by the Generalitat de Catalunya, Departament d’Universitats, Recerca i Societat de la Informació
(2009 SGR 1256), by the Ministerio de Sanidad y Consumo, Instituto de Salud Carlos III, Spanish Network for the Research in Infectious Diseases (REIPI RE06/0008), by the European Community (TROCAR contract HEALTH-F3-2008-223031) and by the Intramural Research Program of the NIDDK, National Institutes of Health. A.F. is sponsored by the Barcelona Institute ADAMTS5 for Global Health (ISGlobal). “
“A protein glycosylation system related to that for protein mannosylation in yeast is present in many actinomycetes. This system involves polyprenyl phosphate mannose synthase (Ppm), protein mannosyl transferase (Pmt), and lipoprotein N-acyl transferase (Lnt). In this study, we obtained a series of mutants in the ppm (sco1423), lnt1 (sco1014), and pmt (sco3154) genes of Streptomyces coelicolor, which encode Ppm, Lnt1, and Pmt, to analyze their requirement for glycosylation of the heterologously expressed Apa glycoprotein of Mycobacterium tuberculosis.