Document Type
Article
Publication Title
Journal of Bacteriology
Publisher
American Society for Microbiology
Publication Date
10-2019
Volume
201
Issue
22
Disciplines
Biology | Life Sciences
Abstract
Coxiella burnetii is an obligate intracellular gammaproteobacterium and zoonotic agent of Q fever. We previously identified 15 small noncoding RNAs (sRNAs) of C. burnetii. One of them, CbsR12 (Coxiella burnetii small RNA 12), is highly transcribed during axenic growth and becomes more prominent during infection of cultured mammalian cells. Secondary structure predictions of CbsR12 revealed four putative CsrA-binding sites in stem loops with consensus AGGA/ANGGA motifs. We subsequently determined that CbsR12 binds to recombinant C. burnetii CsrA-2, but not CsrA-1, proteins in vitro. Moreover, through a combination of in vitro and cell culture assays, we identified several in trans mRNA targets of CbsR12. Of these, we determined that CbsR12 binds and upregulates translation of carA transcripts coding for carbamoyl phosphate synthetase A, an enzyme that catalyzes the first step of pyrimidine biosynthesis. In addition, CbsR12 binds and downregulates translation of metK transcripts coding for S-adenosylmethionine synthetase, a component of the methionine cycle. Furthermore, we found that CbsR12 binds to and downregulates the quantity of cvpD transcripts, coding for a type IVB effector protein, in mammalian cell culture. Finally, we found that CbsR12 is necessary for expansion of Coxiellacontaining vacuoles and affects growth rates in a dose-dependent manner in the early phase of infecting THP-1 cells. This is the first characterization of a trans-acting sRNA of C. burnetii and the first example of a bacterial sRNA that regulates both CarA and MetK synthesis. CbsR12 is one of only a few identified trans-acting sRNAs that interacts with CsrA.
Keywords
AhcY, CarA, Coxiella burnetii, CsrA, CvpD, MetK, noncoding RNA, posttranscriptional regulation, small RNA
DOI
https://doi.org/10.1128/JB .00524-19
Rights
© 2019 American Society for Microbiology. All Rights Reserved.
Recommended Citation
Wachter, Shaun; Bonazzi, Matteo; Shifflett, Kyle; Moses, Abraham S.; Raghavan, Rahul; and Minnick, Michael F., "A CsrA-Binding, trans-Acting sRNA of Coxiella burnetii Is Necessary for Optimal Intracellular Growth and Vacuole Formation during Early Infection of Host Cells" (2019). Biological Sciences Faculty Publications. 450.
https://scholarworks.umt.edu/biosci_pubs/450