Sortase-catalyzed assembly of distinct heteromeric fimbriae in Actinomyces naeslundii

被引:81
作者
Mishra, Arunima
Das, Asis
Cisar, John O.
Ton-That, Hung
机构
[1] Univ Connecticut, Dept Mol Micorbial & Struct Biol, Ctr Hlth, Farmington, CT 06030 USA
[2] NIDCR, Oral Infect & Immun Branch, NIH, Bethesda, MD 20892 USA
关键词
D O I
10.1128/JB.01952-06
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Two types of adhesive fimbriae are expressed by Actinomyces; however, the architecture and the mechanism of assembly of these structures remain poorly understood. In this study we characterized two fimbrial gene clusters present in the genome of Actinomyces naeslundii strain MG-1. By using immunoelectron microscopy and biochemical analysis, we showed that the fimQ-fimP-srtC1-fimR gene cluster encodes a fimbrial structure (designated type 1) that contains a major subunit, FimP, forming the shaft and a minor subunit, FimQ, located primarily at the tip. Similarly, the fimB-fimA-srtC2 gene cluster encodes a distinct fimbrial structure (designated type 2) composed of a shaft protein, FimA, and a tip protein, FimB. By using allelic exchange, we constructed an in-frame deletion mutant that lacks the SrtC2 sortase. This mutant produces abundant type 1 fimbriae and expresses the monomeric FimA and FimB proteins, but it does not assemble type 2 fimbriae. Thus, SrtC2 is a fimbria-specific sortase that is essential for assembly of the type 2 fimbriae. Together, our experiments pave the way for several lines of molecular investigation that are necessary to elucidate the fimbrial assembly pathways in Actinomyces and their function in the pathogenesis of different biofilm-related oral diseases.
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页码:3156 / 3165
页数:10
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