title: Deletion of a 77-base-pair inverted repeat element alters the synthesis of surface polysaccharides in Porphyromonas gingivalis.
abstract: Bacterial cell surface glycans, such as capsular polysaccharides and lipopolysaccharides (LPS), influence host recognition and are considered key virulence determinants. The periodontal pathogen Porphyromonas gingivalis is known to display at least three different types of surface glycans: O-LPS, A-LPS, and K-antigen capsule. We have shown that PG0121 (in strain W83) encodes a DNABII histone-like protein and that this gene is transcriptionally linked to the K-antigen capsule synthesis genes, generating a large ∼19.4-kb transcript (PG0104-PG0121). Furthermore, production of capsule is deficient in a PG0121 mutant strain. In this study, we report on the identification of an antisense RNA (asRNA) molecule located within a 77-bp inverted repeat (77bpIR) element located near the 5' end of the locus. We show that overexpression of this asRNA decreases the amount of capsule produced, indicating that this asRNA can impact capsule synthesis in trans. We also demonstrate that deletion of the 77bpIR element and thereby synthesis of the large 19.4-kb transcript also diminishes, but does not eliminate, capsule synthesis. Surprisingly, LPS structures were also altered by deletion of the 77bpIR element, and reactivity to monoclonal antibodies specific to both O-LPS and A-LPS was eliminated. Additionally, reduced reactivity to these antibodies was also observed in a PG0106 mutant, indicating that this putative glycosyltransferase, which is required for capsule synthesis, is also involved in LPS synthesis in strain W83. We discuss our finding in the context of how DNABII proteins, an antisense RNA molecule, and the 77bpIR element may modulate expression of surface polysaccharides in P. gingivalis.
  
  
  OBJECTIVE
  The periodontal pathogen Porphyromonas gingivalis displays at least three different types of cell surface glycans: O-LPS, A-LPS, and K-antigen capsule. We have shown using Northern analysis that the K-antigen capsule locus encodes a large transcript (∼19.4 kb), encompassing a 77-bp inverted repeat (77bpIR) element near the 5' end. Here, we report on the identification of an antisense RNA (asRNA) encoded within the 77bpIR. We show that overexpression of this asRNA or deletion of the element decreases the amount of capsule. LPS structures were also altered by deletion of the 77bpIR, and reactivity to monoclonal antibodies to both O-LPS and A-LPS was eliminated. Our data indicate that the 77bpIR element is involved in modulating both LPS and capsule synthesis in P. gingivalis.
authors.affiliation: Department of Oral Biology, University of Florida, Gainesville, Florida, USA.
authors.forename: Brian W
authors.initials: BW
authors.lastname: Bainbridge
authors.affiliation: Department of Oral Biology, University of Florida, Gainesville, Florida, USA.
authors.forename: Takanori
authors.initials: T
authors.lastname: Hirano
authors.affiliation: Department of Biological Sciences, University of Idaho, Moscow, Idaho, USA.
authors.forename: Nicole
authors.initials: N
authors.lastname: Grieshaber
authors.affiliation: Department of Oral Biology, University of Florida, Gainesville, Florida, USA mdavey@dental.ufl.edu.
authors.forename: Mary E
authors.initials: ME
authors.lastname: Davey
mesh_terms: D016667:Bacterial Capsules / Q000737:chemistry / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D008070:Lipopolysaccharides / Q000096:biosynthesis; D009154:Mutation; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016966:Porphyromonas gingivalis / Q000378:metabolism*; D015203:Reproducibility of Results; D059467:Transcriptome
chemical_list: D008070:Lipopolysaccharides; D011135:Polysaccharides, Bacterial