In:
PLOS Genetics, Public Library of Science (PLoS), Vol. 19, No. 7 ( 2023-7-10), p. e1010669-
Abstract:
Pathogenic bacteria, such as Yersinia pseudotuberculosis encounter reactive oxygen species (ROS) as one of the first lines of defense in the mammalian host. In return, the bacteria react by mounting an oxidative stress response. Previous global RNA structure probing studies provided evidence for temperature-modulated RNA structures in the 5’-untranslated region (5’-UTR) of various oxidative stress response transcripts, suggesting that opening of these RNA thermometer (RNAT) structures at host-body temperature relieves translational repression. Here, we systematically analyzed the transcriptional and translational regulation of ROS defense genes by RNA-sequencing, qRT-PCR, translational reporter gene fusions, enzymatic RNA structure probing and toeprinting assays. Transcription of four ROS defense genes was upregulated at 37°C. The trxA gene is transcribed into two mRNA isoforms, of which the most abundant short one contains a functional RNAT. Biochemical assays validated temperature-responsive RNAT-like structures in the 5’-UTRs of sodB , sodC and katA . However, they barely conferred translational repression in Y . pseudotuberculosis at 25°C suggesting partially open structures available to the ribosome in the living cell. Around the translation initiation region of katY we discovered a novel, highly efficient RNAT that was primarily responsible for massive induction of KatY at 37°C. By phenotypic characterization of catalase mutants and through fluorometric real-time measurements of the redox-sensitive roGFP2-Orp1 reporter in these strains, we revealed KatA as the primary H 2 O 2 scavenger. Consistent with the upregulation of katY , we observed an improved protection of Y . pseudotuberculosis at 37°C. Our findings suggest a multilayered regulation of the oxidative stress response in Yersinia and an important role of RNAT-controlled katY expression at host body temperature.
Type of Medium:
Online Resource
ISSN:
1553-7404
DOI:
10.1371/journal.pgen.1010669
DOI:
10.1371/journal.pgen.1010669.g001
DOI:
10.1371/journal.pgen.1010669.g002
DOI:
10.1371/journal.pgen.1010669.g003
DOI:
10.1371/journal.pgen.1010669.g004
DOI:
10.1371/journal.pgen.1010669.g005
DOI:
10.1371/journal.pgen.1010669.g006
DOI:
10.1371/journal.pgen.1010669.g007
DOI:
10.1371/journal.pgen.1010669.g008
DOI:
10.1371/journal.pgen.1010669.g009
DOI:
10.1371/journal.pgen.1010669.g010
DOI:
10.1371/journal.pgen.1010669.g011
DOI:
10.1371/journal.pgen.1010669.g012
DOI:
10.1371/journal.pgen.1010669.s001
DOI:
10.1371/journal.pgen.1010669.s002
DOI:
10.1371/journal.pgen.1010669.s003
DOI:
10.1371/journal.pgen.1010669.s004
DOI:
10.1371/journal.pgen.1010669.s005
DOI:
10.1371/journal.pgen.1010669.s006
DOI:
10.1371/journal.pgen.1010669.s007
DOI:
10.1371/journal.pgen.1010669.s008
DOI:
10.1371/journal.pgen.1010669.s009
DOI:
10.1371/journal.pgen.1010669.s010
DOI:
10.1371/journal.pgen.1010669.s011
DOI:
10.1371/journal.pgen.1010669.s012
DOI:
10.1371/journal.pgen.1010669.s013
DOI:
10.1371/journal.pgen.1010669.s014
DOI:
10.1371/journal.pgen.1010669.s015
DOI:
10.1371/journal.pgen.1010669.r001
DOI:
10.1371/journal.pgen.1010669.r002
DOI:
10.1371/journal.pgen.1010669.r003
DOI:
10.1371/journal.pgen.1010669.r004
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2023
detail.hit.zdb_id:
2186725-2
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