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2016 ; 84
(6
): 1775-1784
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Genetic and Molecular Basis of Kingella kingae Encapsulation
#MMPMID27045037
Starr KF
; Porsch EA
; Seed PC
; St Geme JW 3rd
Infect Immun
2016[Jun]; 84
(6
): 1775-1784
PMID27045037
show ga
Kingella kingae is a common cause of invasive disease in young children and was
recently found to produce a polysaccharide capsule containing
N-acetylgalactosamine (GalNAc) and ?-3-deoxy-d-manno-octulosonic acid (?Kdo).
Given the role of capsules as important virulence factors and effective vaccine
antigens, we set out to determine the genetic determinants of K. kingae
encapsulation. Using a transposon library and a screen for nonencapsulated
mutants, we identified the previously identified ctrABCD (ABC transporter)
operon, a lipA (kpsC)-like gene, a lipB (kpsS)-like gene, and a putative
glycosyltransferase gene designated csaA (capsule synthesis type a gene A). These
genes were found to be present at unlinked locations scattered throughout the
genome, an atypical genetic arrangement for Gram-negative bacteria that elaborate
a capsule dependent on an ABC-type transporter for surface localization. The csaA
gene product contains a predicted glycosyltransferase domain with structural
homology to GalNAc transferases and a predicted capsule synthesis domain with
structural homology to Kdo transferases, raising the possibility that this enzyme
is responsible for alternately linking GalNAc to ?Kdo and ?Kdo to GalNAc.
Consistent with this conclusion, mutation of the DXD motif in the GalNAc
transferase domain and of the HP motif in the Kdo transferase domain resulted in
a loss of encapsulation. Examination of intracellular and surface-associated
capsule in deletion mutants and complemented strains further implicated the lipA
(kpsC)-like gene, the lipB (kpsS)-like gene, and the csaA gene in K. kingae
capsule production. These data define the genetic requirements for encapsulation
in K. kingae and demonstrate an atypical organization of capsule synthesis,
assembly, and export genes.
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