Cargando…

Coordinated modulation of multiple processes through phase variation of a c-di-GMP phosphodiesterase in Clostridioides difficile

The opportunistic nosocomial pathogen Clostridioides difficile exhibits phenotypic heterogeneity through phase variation, a stochastic, reversible process that modulates expression. In C. difficile, multiple sequences in the genome undergo inversion through site-specific recombination. Two such loci...

Descripción completa

Detalles Bibliográficos
Autores principales: Reyes Ruiz, Leila M., King, Kathleen A., Agosto-Burgos, Christian, Gamez, Isabella S., Gadda, Nicole C., Garrett, Elizabeth M., Tamayo, Rita
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9286219/
https://www.ncbi.nlm.nih.gov/pubmed/35789350
http://dx.doi.org/10.1371/journal.ppat.1010677
_version_ 1784747958934175744
author Reyes Ruiz, Leila M.
King, Kathleen A.
Agosto-Burgos, Christian
Gamez, Isabella S.
Gadda, Nicole C.
Garrett, Elizabeth M.
Tamayo, Rita
author_facet Reyes Ruiz, Leila M.
King, Kathleen A.
Agosto-Burgos, Christian
Gamez, Isabella S.
Gadda, Nicole C.
Garrett, Elizabeth M.
Tamayo, Rita
author_sort Reyes Ruiz, Leila M.
collection PubMed
description The opportunistic nosocomial pathogen Clostridioides difficile exhibits phenotypic heterogeneity through phase variation, a stochastic, reversible process that modulates expression. In C. difficile, multiple sequences in the genome undergo inversion through site-specific recombination. Two such loci lie upstream of pdcB and pdcC, which encode phosphodiesterases (PDEs) that degrade the signaling molecule c-di-GMP. Numerous phenotypes are influenced by c-di-GMP in C. difficile including cell and colony morphology, motility, colonization, and virulence. In this study, we aimed to assess whether PdcB phase varies, identify the mechanism of regulation, and determine the effects on intracellular c-di-GMP levels and regulated phenotypes. We found that expression of pdcB is heterogeneous and the orientation of the invertible sequence, or ‘pdcB switch’, determines expression. The pdcB switch contains a promoter that when properly oriented promotes pdcB expression. Expression is augmented by an additional promoter upstream of the pdcB switch. Mutation of nucleotides at the site of recombination resulted in phase-locked strains with significant differences in pdcB expression. Characterization of these mutants showed that the pdcB locked-ON mutant has reduced intracellular c-di-GMP compared to the locked-OFF mutant, consistent with increased and decreased PdcB activity, respectively. These alterations in c-di-GMP had concomitant effects on multiple known c-di-GMP regulated processes, indicating that phase variation of PdcB allows C. difficile to coordinately diversify multiple phenotypes in the population to enhance survival.
format Online
Article
Text
id pubmed-9286219
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-92862192022-07-16 Coordinated modulation of multiple processes through phase variation of a c-di-GMP phosphodiesterase in Clostridioides difficile Reyes Ruiz, Leila M. King, Kathleen A. Agosto-Burgos, Christian Gamez, Isabella S. Gadda, Nicole C. Garrett, Elizabeth M. Tamayo, Rita PLoS Pathog Research Article The opportunistic nosocomial pathogen Clostridioides difficile exhibits phenotypic heterogeneity through phase variation, a stochastic, reversible process that modulates expression. In C. difficile, multiple sequences in the genome undergo inversion through site-specific recombination. Two such loci lie upstream of pdcB and pdcC, which encode phosphodiesterases (PDEs) that degrade the signaling molecule c-di-GMP. Numerous phenotypes are influenced by c-di-GMP in C. difficile including cell and colony morphology, motility, colonization, and virulence. In this study, we aimed to assess whether PdcB phase varies, identify the mechanism of regulation, and determine the effects on intracellular c-di-GMP levels and regulated phenotypes. We found that expression of pdcB is heterogeneous and the orientation of the invertible sequence, or ‘pdcB switch’, determines expression. The pdcB switch contains a promoter that when properly oriented promotes pdcB expression. Expression is augmented by an additional promoter upstream of the pdcB switch. Mutation of nucleotides at the site of recombination resulted in phase-locked strains with significant differences in pdcB expression. Characterization of these mutants showed that the pdcB locked-ON mutant has reduced intracellular c-di-GMP compared to the locked-OFF mutant, consistent with increased and decreased PdcB activity, respectively. These alterations in c-di-GMP had concomitant effects on multiple known c-di-GMP regulated processes, indicating that phase variation of PdcB allows C. difficile to coordinately diversify multiple phenotypes in the population to enhance survival. Public Library of Science 2022-07-05 /pmc/articles/PMC9286219/ /pubmed/35789350 http://dx.doi.org/10.1371/journal.ppat.1010677 Text en © 2022 Reyes Ruiz et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Reyes Ruiz, Leila M.
King, Kathleen A.
Agosto-Burgos, Christian
Gamez, Isabella S.
Gadda, Nicole C.
Garrett, Elizabeth M.
Tamayo, Rita
Coordinated modulation of multiple processes through phase variation of a c-di-GMP phosphodiesterase in Clostridioides difficile
title Coordinated modulation of multiple processes through phase variation of a c-di-GMP phosphodiesterase in Clostridioides difficile
title_full Coordinated modulation of multiple processes through phase variation of a c-di-GMP phosphodiesterase in Clostridioides difficile
title_fullStr Coordinated modulation of multiple processes through phase variation of a c-di-GMP phosphodiesterase in Clostridioides difficile
title_full_unstemmed Coordinated modulation of multiple processes through phase variation of a c-di-GMP phosphodiesterase in Clostridioides difficile
title_short Coordinated modulation of multiple processes through phase variation of a c-di-GMP phosphodiesterase in Clostridioides difficile
title_sort coordinated modulation of multiple processes through phase variation of a c-di-gmp phosphodiesterase in clostridioides difficile
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9286219/
https://www.ncbi.nlm.nih.gov/pubmed/35789350
http://dx.doi.org/10.1371/journal.ppat.1010677
work_keys_str_mv AT reyesruizleilam coordinatedmodulationofmultipleprocessesthroughphasevariationofacdigmpphosphodiesteraseinclostridioidesdifficile
AT kingkathleena coordinatedmodulationofmultipleprocessesthroughphasevariationofacdigmpphosphodiesteraseinclostridioidesdifficile
AT agostoburgoschristian coordinatedmodulationofmultipleprocessesthroughphasevariationofacdigmpphosphodiesteraseinclostridioidesdifficile
AT gamezisabellas coordinatedmodulationofmultipleprocessesthroughphasevariationofacdigmpphosphodiesteraseinclostridioidesdifficile
AT gaddanicolec coordinatedmodulationofmultipleprocessesthroughphasevariationofacdigmpphosphodiesteraseinclostridioidesdifficile
AT garrettelizabethm coordinatedmodulationofmultipleprocessesthroughphasevariationofacdigmpphosphodiesteraseinclostridioidesdifficile
AT tamayorita coordinatedmodulationofmultipleprocessesthroughphasevariationofacdigmpphosphodiesteraseinclostridioidesdifficile