The effect of growth temperature on the biosynthesis of Yersinia enterocolitica 0: 3 lipopolysaccharide: temperature regulates the transcription of the rfb but not of the rfa region

Ayman Al-Hendy, Paavo Toivanen, Mikael Skurnik

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

The rfb region of Yersinia enterocolitica O:3 (YeO3) that determines the synthesis of the O-side chain of the lipopolysaccharide was cloned and expressed in Escherichia coli K12 previously. The clone did not show the in vitro temperature variation in O-side chain expression known for YeO3, instead analogous O-side-chain was produced at 25 °C and at 37 °C and both were similar to that produced by YeO3 at 25 °C. In Northern blot analysis marked reduction in the amount of rfb-specific mRNA was observed from YeO3 grown at 37 °C when compared with those grown at 25 °C. On the other hand, equal amounts of rfb-specific mRNA were detected in the E. coli clone at both growth temperatures. This indicates that the transcription of YeO3 rfb region is dramatically repressed at 37 °C. The repressor gene is located outside the rfb region with no analogous locus in E. coli chromosome. We cloned 18.2 kilobase pairs of YeO3 chromosomal DNA that rendered E. coli K12 reactive with 2B5, a YeO3 core-specific monoclonal antibody, in colony blotting, indirect immunofluorescence and immunoblotting. Hence this clone contains the YeO3 rfa region, encompassing the genes involved in the core oligosaccharide biosynthesis. No apparent difference, in the aforementioned tests, was noticed in the expression of the 2B5 epitope at different growth temperatures either in YeO3 or in E. coli. In Northern blot analysis comparable amounts of rfa-specific mRNA were detected at 25 ° and 37 °C. This argues that in YeO3 the core oligosaccharide biosynthesis is not temperature-regulated.

Original languageEnglish (US)
Pages (from-to)81-86
Number of pages6
JournalMicrobial Pathogenesis
Volume10
Issue number1
DOIs
StatePublished - Jan 1 1991

Fingerprint

Yersinia enterocolitica
Lipopolysaccharides
Temperature
Growth
Escherichia coli K12
Clone Cells
Escherichia coli
Oligosaccharides
Northern Blotting
Messenger RNA
Indirect Fluorescent Antibody Technique
Immunoblotting
Genes
Epitopes
Chromosomes
Monoclonal Antibodies

Keywords

  • Yersinia enterocolitica
  • rfa
  • rfb
  • temperature regulation

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

The effect of growth temperature on the biosynthesis of Yersinia enterocolitica 0 : 3 lipopolysaccharide: temperature regulates the transcription of the rfb but not of the rfa region. / Al-Hendy, Ayman; Toivanen, Paavo; Skurnik, Mikael.

In: Microbial Pathogenesis, Vol. 10, No. 1, 01.01.1991, p. 81-86.

Research output: Contribution to journalArticle

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abstract = "The rfb region of Yersinia enterocolitica O:3 (YeO3) that determines the synthesis of the O-side chain of the lipopolysaccharide was cloned and expressed in Escherichia coli K12 previously. The clone did not show the in vitro temperature variation in O-side chain expression known for YeO3, instead analogous O-side-chain was produced at 25 °C and at 37 °C and both were similar to that produced by YeO3 at 25 °C. In Northern blot analysis marked reduction in the amount of rfb-specific mRNA was observed from YeO3 grown at 37 °C when compared with those grown at 25 °C. On the other hand, equal amounts of rfb-specific mRNA were detected in the E. coli clone at both growth temperatures. This indicates that the transcription of YeO3 rfb region is dramatically repressed at 37 °C. The repressor gene is located outside the rfb region with no analogous locus in E. coli chromosome. We cloned 18.2 kilobase pairs of YeO3 chromosomal DNA that rendered E. coli K12 reactive with 2B5, a YeO3 core-specific monoclonal antibody, in colony blotting, indirect immunofluorescence and immunoblotting. Hence this clone contains the YeO3 rfa region, encompassing the genes involved in the core oligosaccharide biosynthesis. No apparent difference, in the aforementioned tests, was noticed in the expression of the 2B5 epitope at different growth temperatures either in YeO3 or in E. coli. In Northern blot analysis comparable amounts of rfa-specific mRNA were detected at 25 ° and 37 °C. This argues that in YeO3 the core oligosaccharide biosynthesis is not temperature-regulated.",
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