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There is growing evidence that GreA aids adaptation to stressful environments in various bacteria. However, the functions of GreA among mycobacteria remain obscure. Here, we report on cellular consequences following deletion of  greA  gene in  Mycobacterium  spp. The  greA  mutant strain (Δ greA ) was generated in  Mycobacterium smegmatis ,  Mycobacterium tuberculosis  (MTB) H37Ra, and  M. tuberculosis  H37Rv. Deletion of  greA  results in growth retardation and poor survival in response to adverse stress, besides rendering  M. tuberculosis  more susceptible to vancomycin and rifampicin. By using RNA-seq, we observe that disrupting  greA  results in the differential regulation of 195 genes in  M. smegmatis  with 167 being negatively regulated. Among these, KEGG pathways significantly enriched for differentially regulated genes included tryptophan metabolism, starch and sucrose metabolism, and carotenoid biosynthesis, supporting a role of GreA in the metabolic regulation of mycobacteria. Moreover, like  Escherichia coli  GreA,  M. smegmatis  GreA exhibits a series of conservative features, and the anti-backtracking activity of C-terminal domain is indispensable for the expression of  glgX , a gene was down-regulated in the RNA-seq data. Interestingly, the decrease in the expression of  glgX  by CRISPR interference, resulted in reduced growth. Finally, intracellular fitness significantly declines due to loss of  greA . Our data indicates that GreA is an important factor for the survival and resistance establishment in  Mycobacterium  spp. This study provides new insight into GreA as a potential target in multi-drug resistant TB treatment.

Involvement of Transcription Elongation Factor GreA in Mycobacterium Viability, Antibiotic Susceptibility, and Intracellular Fitness