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Adhesion to cellulose of the Gram-positive bacterium Ruminococcus albus involves type IV pili

Abstract : This study was aimed at characterizing a cell-surface 25 kDa glycoprotein (GP25) that was previously shown to be underproduced by a spontaneous adhesion-defective mutant D5 of Ruminococcus albus 20. An antiserum against wild-type strain 20 was adsorbed with the mutant D5 to enrich it in antibodies 'specific' to adhesion structures of R. albus 20. The resulting antiserum, called anti-Adh serum, blocked adhesion of R. albus 20 and reacted mainly with GP25 in bacterial and extracellular protein fractions of R. albus 20. The N-terminal sequence of purified GP25 was identical to that of CbpC, a 21 kDa cellulose-binding protein (CBP) of R. albus 8. The nucleotide sequence of the gp25 gene was determined by PCR and genomic walking procedures. The gp25 gene encoded a protein of 165 aa with a calculated molecular mass of 16940 Da that showed 72-9% identity with CbpC and presented homologies with type IV pilins of Gram-negative pathogenic bacteria. Negative-staining electron microscopy revealed fine and flexible pill surrounding R. albus 20 cells while mutant cells were not piliated. In addition, immunoelectron microscopy showed that the anti-Adh serum probing mainly GP25, completely decorated the pill surrounding R. albus 20, thereby showing that GP25 was a major pilus subunit. This study shows for the first time the presence of pill at the surface of R. albus and identifies GP25 as their major protein subunit. Though GP25 was not identified as a CBP, isolated pill were shown to bind cellulose. in conclusion, these pill, which belong to the family of type IV pill, mediate adhesion of R. albus 20 to cellulose.
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https://hal.inrae.fr/hal-02675489
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Submitted on : Sunday, May 31, 2020 - 5:40:40 PM
Last modification on : Tuesday, June 22, 2021 - 3:10:02 PM

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Harivony Rakotoarivonina, Grégory Jubelin, Michel Hébraud, Brigitte Gaillard-Martinie, Evelyne Forano. Adhesion to cellulose of the Gram-positive bacterium Ruminococcus albus involves type IV pili. Microbiology, Microbiology Society, 2002, 148 (6), pp.1871-1880. ⟨hal-02675489⟩

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