Abstract

The RecX protein has attracted considerable interest because the recX mutants exhibit multiple phenotypes associated with RecA functions. To further our understanding of the functional relationship between recA and recX, the effect of different stress treatments on their expression profiles, cell yield and viability were investigated. A significant correlation was found between the expression of Mycobacterium smegmatis recA and recX genes at different stages of growth, and in response to different stress treatments albeit recX exhibiting lower transcript and protein abundance at the mid-log and stationary phases of the bacterial growth cycle. To ascertain their roles in vivo, a targeted deletion of the recX and recArecX was performed in M. smegmatis. The growth kinetics of these mutant strains and their sensitivity patterns to different stress treatments were assessed relative to the wild-type strain. The deletion of recA affected normal cell growth and survival, while recX deletion showed no significant effect. Interestingly, deletion of both recX and recA genes results in a phenotype that is intermediate between the phenotypes of the ΔrecA mutant and the wild-type strain. Collectively, these results reveal a previously unrecognized role for M. smegmatis recX and support the notion that it may regulate a subset of the yet unknown genes involved in normal cell growth and DNA-damage repair.

Highlights

  • To maintain genomic integrity, bacteria have developed a complex network of DNA-damage response (DDR) pathways to sense, respond to, and repair different types of DNA damage[1,2,3]

  • During the course of infection and proliferation, M. tuberculosis is exposed to physiological stress conditions such as hypoxia and acidic pH stress, which could compromise its survival[38,39,40]

  • The recX transcript was hardly detectable during the vegetative growth of E. coli, but robust expression of recX occurs following treatment with DNA damaging agents[15,55]

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Summary

Introduction

Bacteria have developed a complex network of DNA-damage response (DDR) pathways to sense, respond to, and repair different types of DNA damage[1,2,3]. Other known negative modulators include RdgC and PsiB: whereas RdgC acts as a negative regulator of RecA protein functions, PsiB binds to the RecA that is free in solution and the resultant complex impedes the formation of RecA nucleoprotein filaments[16,17,18,19] Another accessory factor, RecN, a bacterial SMC (Structural Maintenance of Chromosome)-like accessory protein, stimulates the DNA strand-invasion step of www.nature.com/scientificreports/. A significant correlation was seen between the expression of the recA and recX at different stages of growth and after exposure to DNA damaging agents, albeit recX exhibiting lower transcript and protein abundance at the mid-log and stationary phases of growth These results suggest a previously unrecognized role for M. smegmatis recX and support the idea that it may regulate a subset of the yet unknown genes involved in normal cell growth and DNA damage repair

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