Bacterial persistence increases as environmental fitness decreases.
Since persister cells cause chronic infections and since Escherichia coli toxin MqsR increases persisters, we used protein engineering to increase the toxicity of MqsR to gain insights into persister cell formation. Through two amino acid replacements that increased the stability of MqsR, toxicity and persistence were increased. A whole-transcriptome study revealed that the MqsR variant increased persistence by repressing genes for acid resistance, multidrug resistance and osmotic resistance. Corroborating these microarray results, deletion of rpoS, as well as the genes that the master stress response regulator RpoS controls, increased persister formation dramatically to the extent that nearly the whole population became persistent. Furthermore, wild-type cells stressed by prior treatment to acid or hydrogen peroxide increased persistence 12 000-fold. Whole-transcriptome analyses of persister cells generated by two different methods (wild-type cells pretreated with hydrogen peroxide and the rpoS deletion) corroborated the importance of suppressing RpoS in persister cell formation. Therefore, the more toxic MqsR increases persistence by decreasing the ability of the cell to respond to antibiotic stress through its RpoS-based regulation of acid resistance, multidrug resistance and osmotic resistance systems.
Departments of Chemical Engineering             Biology, Texas A&M University, College Station, TX 77843-3122, USA.
Seok Hoon
SH
Hong
Xiaoxue
X
Wang
Hazel F
HF
O'Connor
Michael J
MJ
Benedik
Thomas K
TK
Wood
D000143:Acids / Q000633:toxicity; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D046228:Microarray Analysis; D050296:Microbial Viability / Q000187:drug effects; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D016877:Oxidants / Q000633:toxicity; D012808:Sigma Factor / Q000235:genetics
D000143:Acids; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C508571:MqsR protein, E coli; D050505:Mutant Proteins; D016877:Oxidants; D012808:Sigma Factor; C069567:sigma factor KatF protein, Bacteria