title: The highly conserved bacterial RNase YbeY is essential in Vibrio cholerae, playing a critical role in virulence, stress regulation, and RNA processing.
abstract: YbeY, a highly conserved protein, is an RNase in E. coli and plays key roles in both processing of the critical 3' end of 16 S rRNA and in 70 S ribosome quality control under stress. These central roles account for YbeY's inclusion in the postulated minimal bacterial genome. However, YbeY is not essential in E. coli although loss of ybeY severely sensitizes it to multiple physiological stresses. Here, we show that YbeY is an essential endoribonuclease in Vibrio cholerae and is crucial for virulence, stress regulation, RNA processing and ribosome quality control, and is part of a core set of RNases essential in most representative pathogens. To understand its function, we analyzed the rRNA and ribosome profiles of a V. cholerae strain partially depleted for YbeY and other RNase mutants associated with 16 S rRNA processing; our results demonstrate that YbeY is also crucial for 16 S rRNA 3' end maturation in V. cholerae and that its depletion impedes subunit assembly into 70 S ribosomes. YbeY's importance to V. cholerae pathogenesis was demonstrated by the complete loss of mice colonization and biofilm formation, reduced cholera toxin production, and altered expression levels of virulence-associated small RNAs of a V. cholerae strain partially depleted for YbeY. Notably, the ybeY genes of several distantly related pathogens can fully complement an E. coli ΔybeY strain under various stress conditions, demonstrating the high conservation of YbeY's activity in stress regulation. Taken together, this work provides the first comprehensive exploration of YbeY's physiological role in a human pathogen, showing its conserved function across species in essential cellular processes.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Maarten
authors.initials: M
authors.lastname: Vercruysse
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Caroline
authors.initials: C
authors.lastname: Köhrer
authors.affiliation: Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas, United States of America.
authors.forename: Bryan W
authors.initials: BW
authors.lastname: Davies
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Markus F F
authors.initials: MF
authors.lastname: Arnold
authors.affiliation: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachussets, United States of America.
authors.forename: John J
authors.initials: JJ
authors.lastname: Mekalanos
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Uttam L
authors.initials: UL
authors.lastname: RajBhandary
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Graham C
authors.initials: GC
authors.lastname: Walker
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018441:Biofilms / Q000254:growth & development; D002771:Cholera / Q000201:enzymology / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D002772:Cholera Toxin / Q000096:biosynthesis; D017124:Conserved Sequence; D004722:Endoribonucleases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D018928:Immunity, Mucosal; D007413:Intestinal Mucosa / Q000254:growth & development / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D051379:Mice; D009154:Mutation; D010802:Phylogeny; D026721:RNA 3' End Processing; D012329:RNA, Bacterial / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000378:metabolism*; D013312:Stress, Physiological; D014734:Vibrio cholerae / Q000201:enzymology* / Q000276:immunology / Q000472:pathogenicity / Q000502:physiology; D014774:Virulence; D037521:Virulence Factors / Q000096:biosynthesis
chemical_list: D001426:Bacterial Proteins; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D037521:Virulence Factors; D002772:Cholera Toxin; D004722:Endoribonucleases