Advanced

Molecular switch controlling expression of the mannose-specific adhesin, Msa, in Lactobacillus plantarum

Holst, Bjørn; Glenting, Jacob; Holmstrøm, Kim; Israelsen, Hans; Vrang, Astrid; Antonsson, Martin LU ; Ahrné, Siv LU and Madsen, Søren M. (2019) In Applied and Environmental Microbiology 85(10).
Abstract

Some lactic acid bacteria, especially Lactobacillus spp., possess adhesive properties enabling colonization of the human gastrointestinal tract. Two probiotic Lactobacillus plantarum strains, WCSF1 and 299v, display highly different mannose-specific adhesion, with L. plantarum 299v being superior to L. plantarum WCFS1 based on a yeast agglutination assay. A straightforward correlation between the mannose adhesion capacity and domain composition of the mannose-specific adhesin (Msa) in the two strains has not been demonstrated previously. In this study, we analyzed the promoter regions upstream of the msa gene encoding a mannose-specific adhesin in these two strains. The promoter region was mapped by primer extension and DNA sequence... (More)

Some lactic acid bacteria, especially Lactobacillus spp., possess adhesive properties enabling colonization of the human gastrointestinal tract. Two probiotic Lactobacillus plantarum strains, WCSF1 and 299v, display highly different mannose-specific adhesion, with L. plantarum 299v being superior to L. plantarum WCFS1 based on a yeast agglutination assay. A straightforward correlation between the mannose adhesion capacity and domain composition of the mannose-specific adhesin (Msa) in the two strains has not been demonstrated previously. In this study, we analyzed the promoter regions upstream of the msa gene encoding a mannose-specific adhesin in these two strains. The promoter region was mapped by primer extension and DNA sequence analysis, and only a single nucleotide change was identified between the two strains. However, Northern blot analysis showed a stronger msa transcript band in 299v than in WCFS1 correlating with the different adhesion capacities. During the establishment of a high-throughput yeast agglutination assay, we isolated variants of WCFS1 that displayed a very strong mannose-specific adhesion phenotype. The region upstream of the msa gene in these variants showed an inversion of a 104-bp fragment located between two perfectly inverted repeats present in the untranslated leader region. The inversion disrupts a strong hairpin structure that otherwise most likely would terminate the msa transcript. In addition, the ribosome binding site upstream of the msa gene, which is also masked within this hairpin structure, becomes accessible upon inversion, thereby increasing the frequency of translation initiation in the variant strains. Furthermore, Northern blot analysis showed a higher abundance of the msa transcript in the variants than in the wild type, correlating with a strong-Msa phenotype.

(Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Lactobacillus plantarum, Mannose-specific adhesin, Probiotics, Site-specific DNA inversion systems
in
Applied and Environmental Microbiology
volume
85
issue
10
publisher
American Society for Microbiology
external identifiers
  • scopus:85065585655
ISSN
0099-2240
DOI
10.1128/AEM.02954-18
language
English
LU publication?
yes
id
dacb9cd8-9ee2-4177-9640-d69a163118c7
date added to LUP
2019-05-23 12:45:47
date last changed
2019-06-19 04:13:28
@article{dacb9cd8-9ee2-4177-9640-d69a163118c7,
  abstract     = {<p>Some lactic acid bacteria, especially Lactobacillus spp., possess adhesive properties enabling colonization of the human gastrointestinal tract. Two probiotic Lactobacillus plantarum strains, WCSF1 and 299v, display highly different mannose-specific adhesion, with L. plantarum 299v being superior to L. plantarum WCFS1 based on a yeast agglutination assay. A straightforward correlation between the mannose adhesion capacity and domain composition of the mannose-specific adhesin (Msa) in the two strains has not been demonstrated previously. In this study, we analyzed the promoter regions upstream of the msa gene encoding a mannose-specific adhesin in these two strains. The promoter region was mapped by primer extension and DNA sequence analysis, and only a single nucleotide change was identified between the two strains. However, Northern blot analysis showed a stronger msa transcript band in 299v than in WCFS1 correlating with the different adhesion capacities. During the establishment of a high-throughput yeast agglutination assay, we isolated variants of WCFS1 that displayed a very strong mannose-specific adhesion phenotype. The region upstream of the msa gene in these variants showed an inversion of a 104-bp fragment located between two perfectly inverted repeats present in the untranslated leader region. The inversion disrupts a strong hairpin structure that otherwise most likely would terminate the msa transcript. In addition, the ribosome binding site upstream of the msa gene, which is also masked within this hairpin structure, becomes accessible upon inversion, thereby increasing the frequency of translation initiation in the variant strains. Furthermore, Northern blot analysis showed a higher abundance of the msa transcript in the variants than in the wild type, correlating with a strong-Msa phenotype.</p>},
  articleno    = {e02954-18},
  author       = {Holst, Bjørn and Glenting, Jacob and Holmstrøm, Kim and Israelsen, Hans and Vrang, Astrid and Antonsson, Martin and Ahrné, Siv and Madsen, Søren M.},
  issn         = {0099-2240},
  keyword      = {Lactobacillus plantarum,Mannose-specific adhesin,Probiotics,Site-specific DNA inversion systems},
  language     = {eng},
  number       = {10},
  publisher    = {American Society for Microbiology},
  series       = {Applied and Environmental Microbiology},
  title        = {Molecular switch controlling expression of the mannose-specific adhesin, Msa, in Lactobacillus plantarum},
  url          = {http://dx.doi.org/10.1128/AEM.02954-18},
  volume       = {85},
  year         = {2019},
}