Five New Genes Are Important for Common Polysaccharide Antigen Biosynthesis in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>

ABSTRACT Common polysaccharide antigen (CPA) is a conserved cell surface polysaccharide produced by Pseudomonas aeruginosa. It contains a rhamnan homopolymer and is one of the two forms of O polysaccharide attached to P. aeruginosa lipopolysaccharide (LPS). Our laboratory has previously characterize...

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Autores principales:	Youai Hao, Jerry D. King, Steven Huszczynski, Dana Kocíncová, Joseph S. Lam
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Lenguaje:	EN
Publicado:	American Society for Microbiology 2013
Materias:	Microbiology QR1-502
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spelling	oai:doaj.org-article:7cd4b673aa3c4aeda00fa539a1ca1d5b2021-11-15T15:40:23ZFive New Genes Are Important for Common Polysaccharide Antigen Biosynthesis in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>10.1128/mBio.00631-122150-7511https://doaj.org/article/7cd4b673aa3c4aeda00fa539a1ca1d5b2013-03-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00631-12https://doaj.org/toc/2150-7511ABSTRACT Common polysaccharide antigen (CPA) is a conserved cell surface polysaccharide produced by Pseudomonas aeruginosa. It contains a rhamnan homopolymer and is one of the two forms of O polysaccharide attached to P. aeruginosa lipopolysaccharide (LPS). Our laboratory has previously characterized an eight-gene cluster (pa5447-pa5454 in P. aeruginosa PAO1) required for biosynthesis of CPA. Here we demonstrate that an adjacent five-gene cluster pa5455-pa5459 is also involved. Using reverse transcriptase PCR (RT-PCR), we showed that the original eight-gene cluster and the new five-gene cluster are both organized as operons. We have analyzed the LPS phenotypes of in-frame deletion mutants made in each of the five genes, and the results verified that these five genes are indeed required for CPA biosynthesis, extending the CPA biosynthesis locus to contain 13 contiguous genes. By performing overexpression experiments of different sets of these biosynthesis genes, we were able to obtain information about their possible functions in CPA biosynthesis. IMPORTANCE Lipopolysaccharide (LPS) is an important cell surface structure of Gram-negative bacteria. The human opportunistic pathogen Pseudomonas aeruginosa simultaneously produces an O-antigen-specific (OSA) form and a common polysaccharide antigen (CPA) form of LPS. CPA, the focus of this study, is composed of α-1-2, α1-3-linked d-rhamnose sugars and has been shown to be important for attachment of the bacteria to human airway epithelial cells. Genome sequencing of this species revealed a new five-gene cluster that we predicted to be involved in CPA biosynthesis and modification. In this study, we have generated chromosomal knockouts by performing in-frame deletions and allelic replacements. Characterizing the function of each of the five genes is important for us to better understand CPA biosynthesis and the mechanisms of chain length termination and regulation of this unique D-rhamnan polysaccharide.Youai HaoJerry D. KingSteven HuszczynskiDana KocíncováJoseph S. LamAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 4, Iss 1 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Youai Hao Jerry D. King Steven Huszczynski Dana Kocíncová Joseph S. Lam Five New Genes Are Important for Common Polysaccharide Antigen Biosynthesis in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
description	ABSTRACT Common polysaccharide antigen (CPA) is a conserved cell surface polysaccharide produced by Pseudomonas aeruginosa. It contains a rhamnan homopolymer and is one of the two forms of O polysaccharide attached to P. aeruginosa lipopolysaccharide (LPS). Our laboratory has previously characterized an eight-gene cluster (pa5447-pa5454 in P. aeruginosa PAO1) required for biosynthesis of CPA. Here we demonstrate that an adjacent five-gene cluster pa5455-pa5459 is also involved. Using reverse transcriptase PCR (RT-PCR), we showed that the original eight-gene cluster and the new five-gene cluster are both organized as operons. We have analyzed the LPS phenotypes of in-frame deletion mutants made in each of the five genes, and the results verified that these five genes are indeed required for CPA biosynthesis, extending the CPA biosynthesis locus to contain 13 contiguous genes. By performing overexpression experiments of different sets of these biosynthesis genes, we were able to obtain information about their possible functions in CPA biosynthesis. IMPORTANCE Lipopolysaccharide (LPS) is an important cell surface structure of Gram-negative bacteria. The human opportunistic pathogen Pseudomonas aeruginosa simultaneously produces an O-antigen-specific (OSA) form and a common polysaccharide antigen (CPA) form of LPS. CPA, the focus of this study, is composed of α-1-2, α1-3-linked d-rhamnose sugars and has been shown to be important for attachment of the bacteria to human airway epithelial cells. Genome sequencing of this species revealed a new five-gene cluster that we predicted to be involved in CPA biosynthesis and modification. In this study, we have generated chromosomal knockouts by performing in-frame deletions and allelic replacements. Characterizing the function of each of the five genes is important for us to better understand CPA biosynthesis and the mechanisms of chain length termination and regulation of this unique D-rhamnan polysaccharide.
format	article
author	Youai Hao Jerry D. King Steven Huszczynski Dana Kocíncová Joseph S. Lam
author_facet	Youai Hao Jerry D. King Steven Huszczynski Dana Kocíncová Joseph S. Lam
author_sort	Youai Hao
title	Five New Genes Are Important for Common Polysaccharide Antigen Biosynthesis in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_short	Five New Genes Are Important for Common Polysaccharide Antigen Biosynthesis in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_full	Five New Genes Are Important for Common Polysaccharide Antigen Biosynthesis in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_fullStr	Five New Genes Are Important for Common Polysaccharide Antigen Biosynthesis in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_full_unstemmed	Five New Genes Are Important for Common Polysaccharide Antigen Biosynthesis in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_sort	five new genes are important for common polysaccharide antigen biosynthesis in <named-content content-type="genus-species">pseudomonas aeruginosa</named-content>
publisher	American Society for Microbiology
publishDate	2013
url	https://doaj.org/article/7cd4b673aa3c4aeda00fa539a1ca1d5b
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Five New Genes Are Important for Common Polysaccharide Antigen Biosynthesis in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>

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