Cargando…
Discovery of Unannotated Small Open Reading Frames in Streptococcus pneumoniae D39 Involved in Quorum Sensing and Virulence Using Ribosome Profiling
Streptococcus pneumoniae, an opportunistic human pathogen, is the leading cause of community-acquired pneumonia and an agent of otitis media, septicemia, and meningitis. Although genomic and transcriptomic studies of S. pneumoniae have provided detailed perspectives on gene content and expression pr...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9426450/ https://www.ncbi.nlm.nih.gov/pubmed/35852327 http://dx.doi.org/10.1128/mbio.01247-22 |
_version_ | 1784778682188955648 |
---|---|
author | Laczkovich, Irina Mangano, Kyle Shao, Xinhao Hockenberry, Adam J. Gao, Yu Mankin, Alexander Vázquez-Laslop, Nora Federle, Michael J. |
author_facet | Laczkovich, Irina Mangano, Kyle Shao, Xinhao Hockenberry, Adam J. Gao, Yu Mankin, Alexander Vázquez-Laslop, Nora Federle, Michael J. |
author_sort | Laczkovich, Irina |
collection | PubMed |
description | Streptococcus pneumoniae, an opportunistic human pathogen, is the leading cause of community-acquired pneumonia and an agent of otitis media, septicemia, and meningitis. Although genomic and transcriptomic studies of S. pneumoniae have provided detailed perspectives on gene content and expression programs, they have lacked information pertaining to the translational landscape, particularly at a resolution that identifies commonly overlooked small open reading frames (sORFs), whose importance is increasingly realized in metabolism, regulation, and virulence. To identify protein-coding sORFs in S. pneumoniae, antibiotic-enhanced ribosome profiling was conducted. Using translation inhibitors, 114 novel sORFs were detected, and the expression of a subset of them was experimentally validated. Two loci associated with virulence and quorum sensing were examined in deeper detail. One such sORF, rio3, overlaps with the noncoding RNA srf-02 that was previously implicated in pathogenesis. Targeted mutagenesis parsing rio3 from srf-02 revealed that rio3 is responsible for the fitness defect seen in a murine nasopharyngeal colonization model. Additionally, two novel sORFs located adjacent to the quorum sensing receptor rgg1518 were found to impact regulatory activity. Our findings emphasize the importance of sORFs present in the genomes of pathogenic bacteria and underscore the utility of ribosome profiling for identifying the bacterial translatome. |
format | Online Article Text |
id | pubmed-9426450 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-94264502022-08-31 Discovery of Unannotated Small Open Reading Frames in Streptococcus pneumoniae D39 Involved in Quorum Sensing and Virulence Using Ribosome Profiling Laczkovich, Irina Mangano, Kyle Shao, Xinhao Hockenberry, Adam J. Gao, Yu Mankin, Alexander Vázquez-Laslop, Nora Federle, Michael J. mBio Research Article Streptococcus pneumoniae, an opportunistic human pathogen, is the leading cause of community-acquired pneumonia and an agent of otitis media, septicemia, and meningitis. Although genomic and transcriptomic studies of S. pneumoniae have provided detailed perspectives on gene content and expression programs, they have lacked information pertaining to the translational landscape, particularly at a resolution that identifies commonly overlooked small open reading frames (sORFs), whose importance is increasingly realized in metabolism, regulation, and virulence. To identify protein-coding sORFs in S. pneumoniae, antibiotic-enhanced ribosome profiling was conducted. Using translation inhibitors, 114 novel sORFs were detected, and the expression of a subset of them was experimentally validated. Two loci associated with virulence and quorum sensing were examined in deeper detail. One such sORF, rio3, overlaps with the noncoding RNA srf-02 that was previously implicated in pathogenesis. Targeted mutagenesis parsing rio3 from srf-02 revealed that rio3 is responsible for the fitness defect seen in a murine nasopharyngeal colonization model. Additionally, two novel sORFs located adjacent to the quorum sensing receptor rgg1518 were found to impact regulatory activity. Our findings emphasize the importance of sORFs present in the genomes of pathogenic bacteria and underscore the utility of ribosome profiling for identifying the bacterial translatome. American Society for Microbiology 2022-07-19 /pmc/articles/PMC9426450/ /pubmed/35852327 http://dx.doi.org/10.1128/mbio.01247-22 Text en Copyright © 2022 Laczkovich et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Article Laczkovich, Irina Mangano, Kyle Shao, Xinhao Hockenberry, Adam J. Gao, Yu Mankin, Alexander Vázquez-Laslop, Nora Federle, Michael J. Discovery of Unannotated Small Open Reading Frames in Streptococcus pneumoniae D39 Involved in Quorum Sensing and Virulence Using Ribosome Profiling |
title | Discovery of Unannotated Small Open Reading Frames in Streptococcus pneumoniae D39 Involved in Quorum Sensing and Virulence Using Ribosome Profiling |
title_full | Discovery of Unannotated Small Open Reading Frames in Streptococcus pneumoniae D39 Involved in Quorum Sensing and Virulence Using Ribosome Profiling |
title_fullStr | Discovery of Unannotated Small Open Reading Frames in Streptococcus pneumoniae D39 Involved in Quorum Sensing and Virulence Using Ribosome Profiling |
title_full_unstemmed | Discovery of Unannotated Small Open Reading Frames in Streptococcus pneumoniae D39 Involved in Quorum Sensing and Virulence Using Ribosome Profiling |
title_short | Discovery of Unannotated Small Open Reading Frames in Streptococcus pneumoniae D39 Involved in Quorum Sensing and Virulence Using Ribosome Profiling |
title_sort | discovery of unannotated small open reading frames in streptococcus pneumoniae d39 involved in quorum sensing and virulence using ribosome profiling |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9426450/ https://www.ncbi.nlm.nih.gov/pubmed/35852327 http://dx.doi.org/10.1128/mbio.01247-22 |
work_keys_str_mv | AT laczkovichirina discoveryofunannotatedsmallopenreadingframesinstreptococcuspneumoniaed39involvedinquorumsensingandvirulenceusingribosomeprofiling AT manganokyle discoveryofunannotatedsmallopenreadingframesinstreptococcuspneumoniaed39involvedinquorumsensingandvirulenceusingribosomeprofiling AT shaoxinhao discoveryofunannotatedsmallopenreadingframesinstreptococcuspneumoniaed39involvedinquorumsensingandvirulenceusingribosomeprofiling AT hockenberryadamj discoveryofunannotatedsmallopenreadingframesinstreptococcuspneumoniaed39involvedinquorumsensingandvirulenceusingribosomeprofiling AT gaoyu discoveryofunannotatedsmallopenreadingframesinstreptococcuspneumoniaed39involvedinquorumsensingandvirulenceusingribosomeprofiling AT mankinalexander discoveryofunannotatedsmallopenreadingframesinstreptococcuspneumoniaed39involvedinquorumsensingandvirulenceusingribosomeprofiling AT vazquezlaslopnora discoveryofunannotatedsmallopenreadingframesinstreptococcuspneumoniaed39involvedinquorumsensingandvirulenceusingribosomeprofiling AT federlemichaelj discoveryofunannotatedsmallopenreadingframesinstreptococcuspneumoniaed39involvedinquorumsensingandvirulenceusingribosomeprofiling |