A Chlamydia trachomatis strain with a chemically generated amino acid substitution (P370L) in the cthtrA gene shows reduced elementary body production. Issue 1 (December 2015)

• Record Type: Journal Article
• Authors: Marsh, James; Wee, Bryan; Tyndall, Joel; Lott, William; Bastidas, Robert; Caldwell, Harlan; Valdivia, Raphael; Kari, L.; Huston, Wilhelmina
• Is Part Of: BMC microbiology. Volume 15:Issue 1(2015)
• Page Start: 1
• Page End: 13
• Publication Date: 2015-12
• Subjects: Chlamydia -- HtrA -- Genetics -- Heat shock -- Intracellular; Microbiology -- Periodicals; 579.05
• DOI: 10.1186/s12866-015-0533-2
• Languages: English
• ISSNs: 1471-2180

Abstract

Abstract Background Chlamydia (C. )trachomatis is the most prevalent bacterial sexually transmitted infection worldwide and the leading cause of preventable blindness. Genetic approaches to investigateC. trachomatis have been only recently developed due to the organism's intracellular developmental cycle. HtrA is a critical stress response serine protease and chaperone for many bacteria and inC. trachomatis has been previously shown to be important for heat stress and the replicative phase of development using a chemical inhibitor of theCt HtrA activity. In this study, chemically-induced SNVs in thecthtrA gene that resulted in amino acid substitutions (A240V, G475E, and P370L) were identified and characterized. Methods SNVs were initially biochemically characterized in vitro using recombinant protein techniques to confirm a functional impact on proteolysis. TheC. trachomatis strains containing the SNVs with marked reductions in proteolysis were investigated in cell culture to identify phenotypes that could be linked toCt HtrA function. Results The strain harboring the SNV with the most marked impact on proteolysis (cthtrA P370L ) was detected to have a significant reduction in the production of infectious elementary bodies. Conclusions This provides genetic evidence thatCt HtrA is critical for theC. trachomatis developmental cycle.