Bacteroides expand the functional versatility of a conserved transcription factor and transcribed DNA to program capsule diversity

Abstract The genomes of human gut bacteria in the genus Bacteroides include numerous operons for biosynthesis of diverse capsular polysaccharides (CPSs). The first two genes of each CPS operon encode a locus-specific paralog of transcription elongation factor NusG (called UpxY), which enhances trans...

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Main Authors: Jason Saba, Katia Flores, Bailey Marshall, Michael D. Engstrom, Yikai Peng, Atharv S. Garje, Laurie E. Comstock, Robert Landick
Format: Article
Language:English
Published: Nature Portfolio 2024-12-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-024-55215-9
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author Jason Saba
Katia Flores
Bailey Marshall
Michael D. Engstrom
Yikai Peng
Atharv S. Garje
Laurie E. Comstock
Robert Landick
author_facet Jason Saba
Katia Flores
Bailey Marshall
Michael D. Engstrom
Yikai Peng
Atharv S. Garje
Laurie E. Comstock
Robert Landick
author_sort Jason Saba
collection DOAJ
description Abstract The genomes of human gut bacteria in the genus Bacteroides include numerous operons for biosynthesis of diverse capsular polysaccharides (CPSs). The first two genes of each CPS operon encode a locus-specific paralog of transcription elongation factor NusG (called UpxY), which enhances transcript elongation, and a UpxZ protein that inhibits noncognate UpxYs. This process, together with promoter inversions, ensures that a single CPS operon is transcribed in most cells. Here, we use in-vivo nascent-RNA sequencing and promoter-less in-vitro transcription (PIVoT) to show that UpxY recognizes a paused RNA polymerase via sequences in both the exposed non-template DNA and the upstream duplex DNA. UpxY association is aided by ‘pause-then-escape’ nascent RNA hairpins. UpxZ binds non-cognate UpxYs to directly inhibit UpxY association. This UpxY-UpxZ hierarchical regulatory program allows Bacteroides to generate subpopulations of cells producing diverse CPSs for optimal fitness.
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spelling doaj-art-2d7443121d8c44f2a8928db149361fcd2025-08-20T01:48:08ZengNature PortfolioNature Communications2041-17232024-12-0115111610.1038/s41467-024-55215-9Bacteroides expand the functional versatility of a conserved transcription factor and transcribed DNA to program capsule diversityJason Saba0Katia Flores1Bailey Marshall2Michael D. Engstrom3Yikai Peng4Atharv S. Garje5Laurie E. Comstock6Robert Landick7Department of Biochemistry, University of Wisconsin–MadisonDepartment of Microbiology, University of ChicagoDepartment of Biochemistry, University of Wisconsin–MadisonDepartment of Biochemistry, University of Wisconsin–MadisonDepartment of Biochemistry, University of Wisconsin–MadisonDepartment of Biochemistry, University of Wisconsin–MadisonDepartment of Microbiology, University of ChicagoDepartment of Biochemistry, University of Wisconsin–MadisonAbstract The genomes of human gut bacteria in the genus Bacteroides include numerous operons for biosynthesis of diverse capsular polysaccharides (CPSs). The first two genes of each CPS operon encode a locus-specific paralog of transcription elongation factor NusG (called UpxY), which enhances transcript elongation, and a UpxZ protein that inhibits noncognate UpxYs. This process, together with promoter inversions, ensures that a single CPS operon is transcribed in most cells. Here, we use in-vivo nascent-RNA sequencing and promoter-less in-vitro transcription (PIVoT) to show that UpxY recognizes a paused RNA polymerase via sequences in both the exposed non-template DNA and the upstream duplex DNA. UpxY association is aided by ‘pause-then-escape’ nascent RNA hairpins. UpxZ binds non-cognate UpxYs to directly inhibit UpxY association. This UpxY-UpxZ hierarchical regulatory program allows Bacteroides to generate subpopulations of cells producing diverse CPSs for optimal fitness.https://doi.org/10.1038/s41467-024-55215-9
spellingShingle Jason Saba
Katia Flores
Bailey Marshall
Michael D. Engstrom
Yikai Peng
Atharv S. Garje
Laurie E. Comstock
Robert Landick
Bacteroides expand the functional versatility of a conserved transcription factor and transcribed DNA to program capsule diversity
Nature Communications
title Bacteroides expand the functional versatility of a conserved transcription factor and transcribed DNA to program capsule diversity
title_full Bacteroides expand the functional versatility of a conserved transcription factor and transcribed DNA to program capsule diversity
title_fullStr Bacteroides expand the functional versatility of a conserved transcription factor and transcribed DNA to program capsule diversity
title_full_unstemmed Bacteroides expand the functional versatility of a conserved transcription factor and transcribed DNA to program capsule diversity
title_short Bacteroides expand the functional versatility of a conserved transcription factor and transcribed DNA to program capsule diversity
title_sort bacteroides expand the functional versatility of a conserved transcription factor and transcribed dna to program capsule diversity
url https://doi.org/10.1038/s41467-024-55215-9
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