The S-layer protein of a Clostridium difficile SLCT-11 strain displays a complex glycan required for normal cell growth and morphology
© 2018 Richards et al..
Clostridium difficile is a bacterial pathogen that causes major health challenges worldwide. It has a well-characterized surface (S)-layer, a para-crystalline proteinaceous layer surrounding the cell wall. In many bacterial and archaeal species, the S-layer is glycosylated, but no such modifications have been demonstrated in C. difficile. Here, we show that a C. difficile strain of S-layer cassette type 11, Ox247, has a complex glycan attached via an O-linkage to Thr-38 of the S-layer low-molecular-weight subunit. Using MS and NMR, we fully characterized this glycan. We present evidence that it is composed of three domains: (i) a core peptide-linked tetrasaccharide with the sequence -4-α-Rha-3-α-Rha-3-α-Rha-3-β-Gal-peptide; (ii) a repeating pentasaccharide with the sequence -4-β-Rha-4-α-Glc-3-β-Rha-4-(α-Rib-3-)β-Rha-; and (iii) a nonreducing end-terminal 2,3 cyclophosphoryl-rhamnose attached to a ribose-branched sub-terminal rhamnose residue. The Ox247 genome contains a 24-kb locus containing genes for synthesis and protein attachment of this glycan. Mutations in genes within this locus altered or completely abrogated formation of this glycan, and their phenotypes suggested that this S-layer modification may affect sporulation, cell length, and biofilm formation of C. difficile In summary, our findings indicate that the S-layer protein of SLCT-11 strains displays a complex glycan and suggest that this glycan is required for C. difficile sporulation and control of cell shape, a discovery with implications for the development of antimicrobials targeting the S-layer.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2018 |
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| Erschienen: |
2018 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:293 |
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| Enthalten in: |
The Journal of biological chemistry - 293(2018), 47 vom: 23. Nov., Seite 18123-18137 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Richards, Emma [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 24.06.2019 Date Revised 04.04.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1074/jbc.RA118.004530 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM289106753 |
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| 100 | 1 | |a Richards, Emma |e verfasserin |4 aut | |
| 245 | 1 | 4 | |a The S-layer protein of a Clostridium difficile SLCT-11 strain displays a complex glycan required for normal cell growth and morphology |
| 264 | 1 | |c 2018 | |
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| 500 | |a Date Completed 24.06.2019 | ||
| 500 | |a Date Revised 04.04.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2018 Richards et al. | ||
| 520 | |a Clostridium difficile is a bacterial pathogen that causes major health challenges worldwide. It has a well-characterized surface (S)-layer, a para-crystalline proteinaceous layer surrounding the cell wall. In many bacterial and archaeal species, the S-layer is glycosylated, but no such modifications have been demonstrated in C. difficile. Here, we show that a C. difficile strain of S-layer cassette type 11, Ox247, has a complex glycan attached via an O-linkage to Thr-38 of the S-layer low-molecular-weight subunit. Using MS and NMR, we fully characterized this glycan. We present evidence that it is composed of three domains: (i) a core peptide-linked tetrasaccharide with the sequence -4-α-Rha-3-α-Rha-3-α-Rha-3-β-Gal-peptide; (ii) a repeating pentasaccharide with the sequence -4-β-Rha-4-α-Glc-3-β-Rha-4-(α-Rib-3-)β-Rha-; and (iii) a nonreducing end-terminal 2,3 cyclophosphoryl-rhamnose attached to a ribose-branched sub-terminal rhamnose residue. The Ox247 genome contains a 24-kb locus containing genes for synthesis and protein attachment of this glycan. Mutations in genes within this locus altered or completely abrogated formation of this glycan, and their phenotypes suggested that this S-layer modification may affect sporulation, cell length, and biofilm formation of C. difficile In summary, our findings indicate that the S-layer protein of SLCT-11 strains displays a complex glycan and suggest that this glycan is required for C. difficile sporulation and control of cell shape, a discovery with implications for the development of antimicrobials targeting the S-layer | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Clostridium difficile | |
| 650 | 4 | |a cell wall | |
| 650 | 4 | |a genetics | |
| 650 | 4 | |a glycomics | |
| 650 | 4 | |a glycoprotein structure | |
| 650 | 4 | |a glycoproteomics | |
| 650 | 4 | |a glycosylation | |
| 650 | 4 | |a mass spectrometry (MS) | |
| 650 | 4 | |a microbiology | |
| 650 | 4 | |a nuclear magnetic resonance (NMR) | |
| 650 | 4 | |a surface layer | |
| 650 | 4 | |a virulence | |
| 650 | 7 | |a Membrane Glycoproteins |2 NLM | |
| 650 | 7 | |a Polysaccharides |2 NLM | |
| 650 | 7 | |a S-layer proteins |2 NLM | |
| 700 | 1 | |a Bouché, Laura |e verfasserin |4 aut | |
| 700 | 1 | |a Panico, Maria |e verfasserin |4 aut | |
| 700 | 1 | |a Arbeloa, Ana |e verfasserin |4 aut | |
| 700 | 1 | |a Vinogradov, Evgeny |e verfasserin |4 aut | |
| 700 | 1 | |a Morris, Howard |e verfasserin |4 aut | |
| 700 | 1 | |a Wren, Brendan |e verfasserin |4 aut | |
| 700 | 1 | |a Logan, Susan M |e verfasserin |4 aut | |
| 700 | 1 | |a Dell, Anne |e verfasserin |4 aut | |
| 700 | 1 | |a Fairweather, Neil F |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t The Journal of biological chemistry |d 1945 |g 293(2018), 47 vom: 23. Nov., Seite 18123-18137 |w (DE-627)NLM000004995 |x 1083-351X |7 nnns |
| 773 | 1 | 8 | |g volume:293 |g year:2018 |g number:47 |g day:23 |g month:11 |g pages:18123-18137 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1074/jbc.RA118.004530 |3 Volltext |
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