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PMID: 10913080 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The serotype of type Ia and III group B streptococci is determined by the polymerase gene within the polycistronic capsule operon.

Journal of bacteriology ·Vol. 182 ·No. 16 ·2000-08-00 ·Pages 4466-77

Chaffin DO, Beres SB, Yim HH, Rubens CE

Abstract

Streptococcus agalactiae is a primary cause of neonatal morbidity and mortality. Essential to the virulence of this pathogen is the production of a type-specific capsular polysaccharide (CPS) that enables the bacteria to evade host immune defenses. The identification, cloning, sequencing, and functional characterization of seven genes involved in type III capsule production have been previously reported. Here, we describe the cloning and sequencing of nine additional adjacent genes, cps(III)FGHIJKL, neu(III)B, and neu(III)C. Sequence comparisons suggested that these genes are involved in sialic acid synthesis, pentasaccharide repeating unit formation, and oligosaccharide transport and polymerization. The type III CPS (cpsIII) locus was comprised of 16 genes within 15.5 kb of contiguous chromosomal DNA. Primer extension analysis and investigation of mRNA from mutants with polar insertions in their cpsIII loci supported the hypothesis that the operon is transcribed as a single polycistronic message. The translated cpsIII sequences were compared to those of the S. agalactiae cpsIa locus, and the primary difference between the operons was found to reside in cps(III)H, the putative CPS polymerase gene. Expression of cps(III)H in a type Ia strain resulted in suppression of CPS Ia synthesis and in production of a CPS which reacted with type III-specific polyclonal antibody. Likewise, expression of the putative type Ia polymerase gene in a type III strain reduced synthesis of type III CPS with production of a type Ia immunoreactive capsule. Based on the similar structures of the oligosaccharide repeating units of the type Ia and III capsules, our observations demonstrated that cps(Ia)H and cps(III)H encoded the type Ia and III CPS polymerases, respectively. Additionally, these findings suggested that a single gene can confer serotype specificity in organisms that produce complex polysaccharides.

MeSH Terms
Base Sequence Carbohydrate Conformation Carbohydrate Sequence Chromosome Mapping Cloning, Molecular Databases, Factual Escherichia coli Introns Molecular Sequence Data N-Acetylneuraminic Acid/biosynthesis Oligosaccharides/chemical synthesis,chemistry Operon Plasmids Polysaccharides, Bacterial/biosynthesis,chemistry,genetics RNA, Messenger/genetics Serotyping Streptococcus agalactiae/classification,enzymology,genetics
Chemicals
Oligosaccharides Polysaccharides, Bacterial RNA, Messenger N-Acetylneuraminic Acid
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Chaffin D O
Department of Pediatrics, Division of Infectious Diseases, Children's Hospital Regional Medical Center, Seattle, Washington 98105, USA.
Beres S B
Yim H H
Rubens C E
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.182.16.4466-4477.2000
Published
2000-08-00
Pages
4466-77
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC94618
Subset
IM
Grants
NIAID NIH HHS · R01 AI022498 · United States
NIAID NIH HHS · AI22152 · United States
NIAID NIH HHS · AI22498 · United States
External Links
PubMed source DOI: 10.1128/JB.182.16.4466-4477.2000 PMC Full Text View journal details
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