Molecular characterization of type-specific capsular polysaccharide biosynthesis genes of Streptococcus agalactiae type Ia

Shin Yamamoto, Katsuhide Miyake, Yoichi Koike, Masaki Watanabe, Yuichi Machida, Michio Ohta, Shinji Iijima

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

The type-specific capsular polysaccharide (CP) of a group B streptococcus, Streptococcus agalactiae type Ia, is a high-molecular-weight polymer consisting of the pentasaccharide repeating unit 4)-[α-D-NeupNAc- (2→3)β-D-Galp-(1→4)-β-D-GlcpNAc-(1→3)]-[β-D-Galp-(1→4)-β-D-Glcp-(1. Here, cloning, sequencing, and transcription of the type Ia-specific capsular polysaccharide synthesis (cps) genes and functional analysis of these gene products are described. A 26-kb DNA fragment containing 18 complete open reading frames (ORFs) was cloned. These ORFs were designated cpsIaA to cpsIaL, neu (neuraminic acid synthesis gene) A to D, orf1 and ung (uracil DNA glycosylase). The cps gene products of S. agalactiae type Ia were homologous to proteins involved in CP synthesis of S. agalactiae type III and S. pneumoniae serotype 14. Unlike the cps gene cluster of S. pneumoniae serotype 14, transcription of this operon may start from cpsIaA, cpsIaE, and orf1 because putative promoter sequences were found in front of these genes. Northern hybridization, reverse transcription-PCR, and primer extension analyses supported this hypothesis. DNA sequence analysis showed that there were two transcriptional terminators in the 3' end of this operon (downstream of orf1 and ung). The functions of CpsIaE, CpsIaG, CpsIaI, and CpsIaJ were examined by glycosyltransferase assay by using the gene products expressed in Escherichia coli JM109 harboring plasmids containing various S. agalactiae type Ia cps gene fragments. Enzyme assays suggested that the gene products of cpsIaE, cpsIaG, cpsIaI, and cpsIaJ are putative glucosyltransferase, β-1,4- galactosyltransferase, β-1,3-N-acetylglucosaminyltransferase, and β-1,4- galactosyltransferase, respectively.

Original languageEnglish (US)
Pages (from-to)5176-5184
Number of pages9
JournalJournal of Bacteriology
Volume181
Issue number17
StatePublished - Sep 1999
Externally publishedYes

Fingerprint

Streptococcus agalactiae
Polysaccharides
Genes
Uracil-DNA Glycosidase
Galactosyltransferases
Operon
Open Reading Frames
Pneumonia
Neuraminic Acids
Glucosyltransferases
Glycosyltransferases
Enzyme Assays
Multigene Family
DNA Sequence Analysis
Reverse Transcription
Organism Cloning
Polymers
Plasmids
Molecular Weight
Escherichia coli

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Yamamoto, S., Miyake, K., Koike, Y., Watanabe, M., Machida, Y., Ohta, M., & Iijima, S. (1999). Molecular characterization of type-specific capsular polysaccharide biosynthesis genes of Streptococcus agalactiae type Ia. Journal of Bacteriology, 181(17), 5176-5184.

Molecular characterization of type-specific capsular polysaccharide biosynthesis genes of Streptococcus agalactiae type Ia. / Yamamoto, Shin; Miyake, Katsuhide; Koike, Yoichi; Watanabe, Masaki; Machida, Yuichi; Ohta, Michio; Iijima, Shinji.

In: Journal of Bacteriology, Vol. 181, No. 17, 09.1999, p. 5176-5184.

Research output: Contribution to journalArticle

Yamamoto, S, Miyake, K, Koike, Y, Watanabe, M, Machida, Y, Ohta, M & Iijima, S 1999, 'Molecular characterization of type-specific capsular polysaccharide biosynthesis genes of Streptococcus agalactiae type Ia', Journal of Bacteriology, vol. 181, no. 17, pp. 5176-5184.
Yamamoto, Shin ; Miyake, Katsuhide ; Koike, Yoichi ; Watanabe, Masaki ; Machida, Yuichi ; Ohta, Michio ; Iijima, Shinji. / Molecular characterization of type-specific capsular polysaccharide biosynthesis genes of Streptococcus agalactiae type Ia. In: Journal of Bacteriology. 1999 ; Vol. 181, No. 17. pp. 5176-5184.
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