Investigating the role of secA2 in secretion and glycosylation of a fimbrial adhesin in Streptococcus parasanguis FW213.

Adhesion of Streptococcus parasanguis FW213, a primary colonizer, to the tooth surface is mediated mainly by peritrichous long fimbriae. The fimbrial structural unit, Fap1, is indispensable for fimbriae biogenesis, adhesion to an in vitro tooth model and biofilm formation. Mature Fap1 is a glycoprotein with an apparent molecular mass of 200 kDa. Glycosylated Fap1 is not present in some mutants screened from a transposon mutant library. Localization of the transposition sites revealed a gene determined to be secA2, which is distinct from the canonical secA gene. In FW213, glycosylated Fap1 was present in all the subcellular fractions including the cytoplasm. In VT1574, a non-polar mutant of secA2 generated by in frame deletion, Fap1 was not secreted. Glycosylated Fap1 was present in the membrane and cytoplasm of the mutant, although in greatly reduced amounts. Fap1 secretion and abundance were restored when VT1574 was complemented by a plasmid-borne secA2. The secretion defect of the secA2 mutation appears to be limited to a small group of proteins such as Fap1 and FimA. These data suggested that Fap1 secretion rather than glycosylation was the major effect of the deletion of secA2; however, this deletion also had an impact on Fap1 abundance. Two more secA2 mutants with different regions deleted were tested for their ability to secrete Fap1. One mutant was completely unable to secrete Fap1 while the other was able to secrete, but in a decreased amount. These data suggest that the region deleted in the latter mutant (nucleotides 2032-2337) is dispensable for Fap1 secretion. Copyright 2004 Blackwell Publishing Ltd