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Infect. Immun., Dec 1996, 5117-5128, Vol 64, No. 12
MM Vickerman, MC Sulavik, PE Minick and DB Clewell
Glucans produced by the glucosyltransferase (GTF) of Streptococcus gordonii
confer a hard, cohesive phenotype (Spp+) on colonies grown on sucrose agar
plates. S. gordonii strains with specific mutations in the region of gtfG
that encodes the GTF carboxyl terminus were characterized. In the parental
strain Challis CH1, this region included a series of six direct repeats
thought to function in glucan binding. The spontaneous mutant strain CH107
had a 585-bp deletion resulting in the loss of three internal direct
repeats. Insertional mutagenesis was used to construct strain CH2RPE, which
had the parental repeat region but was missing 14 carboxyl-terminal amino
acids. The similarly constructed strain CH4RPE had an in-frame addition of
390 nucleotides encoding two additional direct repeats. Although strains
CH1, CH2RPE, and CH4RPE all had similar levels of extracellular GTF
activity, strain CH107 had less than 15% of the parental activity; however,
Western blots (immunoblots) indicated that the amounts of extracellular GTF
protein in all four strains were similar. 13C NMR analyses indicated that
partially purified GTFs from the Spp+ strains CH1, CH2RPE, and CH4RPE all
produced glucans with similar ratios of alpha1,6 and alpha1,3 glucosidic
linkages, whereas the Spp- strain CH107 GTF produced primarily
alpha1,6-linked glucans. Transformation of strain CH107 with pAMS57, which
carries the gtfG positive regulatory determinant, rgg, increased the amount
of GTF activity and GTF antibody- reactive protein ca. fivefold but did not
confer a hard colony phenotype on sucrose agar plates, suggesting that the
type of glucan product affects the sucrose-promoted colony phenotype.
Copyright © 1996, American Society for Microbiology
Changes in the carboxyl-terminal repeat region affect extracellular activity and glucan products of Streptococcus gordonii glucosyltransferase
Department of Microbiology and Immunology, School of Medicine, University of Michigan, Ann Arbor 48109, USA. vickerma@umich.edu
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