Mode of expression and functional characterization of FCT-3 pilus region-encoded proteins in Streptococcus pyogenes serotype M49.
The human pathogen Streptococcus pyogenes (group A streptococcus [GAS]) pilus components, suggested to play a role in pathogenesis, are encoded in the variable FCT (fibronectin- and collagen-binding T-antigen) region. We investigated the functions of sortase A (SrtA), sortase C2 (SrtC2), and the FctA protein of the most prevalent type 3 FCT region from a serotype M49 strain. Although it is considered a housekeeping sortase, SrtA's activity is involved in pilus formation in addition to its essentiality for GAS extracellular matrix protein binding, host cell adherence/internalization, survival in human blood, and biofilm formation. SrtC2 activity is crucial for pilus formation but dispensable for the other phenotypes tested in vitro. FctA is the major pilus backbone protein, simultaneously acting as the M49 T antigen, and requires SrtC2 and LepA, a signal peptidase I homologue, for monomeric surface expression and polymerization, respectively. Collagen-binding protein Cpa expression supports pilus formation at the pilus base. Immunofluorescence microscopy and fluorescence-activated cell sorting analysis revealed several unexpected expression patterns, as follows: (i) the monomeric pilus protein FctA was found exclusively at the old poles of GAS cells, (ii) FctA protein expression increased with lower temperatures, and (iii) FctA protein expression was restricted to 20 to 50% of a given GAS M49 population, suggesting regulation by a bistability mode. Notably, disruption of pilus assembly by sortase deletion rendered GAS serotype M49 significantly more aggressive in a dermonecrotic mouse infection model, indicating that sortase activity and, consequently, pilus expression allow a subpopulation of this GAS serotype to be less aggressive. Thus, pilus expression may not be a virulence attribute of GAS per se.
Institute of Medical Microbiology, Virology and Hygiene, Schillingallee 70, 18055 Rostock, Germany.
Masanobu
M
Nakata
Thomas
T
Köller
Karin
K
Moritz
Deborah
D
Ribardo
Ludwig
L
Jonas
Kevin S
KS
McIver
Tomoko
T
Sumitomo
Yutaka
Y
Terao
Shigetada
S
Kawabata
Andreas
A
Podbielski
Bernd
B
Kreikemeyer
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D001769:Blood / Q000382:microbiology; D002448:Cell Adhesion; D002460:Cell Line; D010861:Fimbriae, Bacterial / Q000235:genetics / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial; D023061:Gene Order; D006801:Humans; D051379:Mice; D050296:Microbial Viability; D012867:Skin / Q000473:pathology; D017192:Skin Diseases, Bacterial / Q000382:microbiology; D013290:Streptococcal Infections / Q000382:microbiology; D013297:Streptococcus pyogenes / Q000235:genetics / Q000502:physiology*; D014774:Virulence; D037521:Virulence Factors / Q000096:biosynthesis* / Q000235:genetics
D001426:Bacterial Proteins; D037521:Virulence Factors