Deletions in the carboxyl-terminal region of Streptococcus gordonii glucosyltransferase affect cell-associated enzyme activity and sucrose-associated accumulation of growing cells.
The single glucosyltransferase (GTF) of Streptococcus gordonii Challis CH1 makes alpha 1,3- and alpha 1,6-linked glucans from sucrose. The GTF carboxyl-terminal region has six direct repeats thought to be involved in glucan binding. Strains with defined mutations in this region have been described recently (M. M. Vickerman, M. C. Sulavik, P. E. Minick, and D. B. Clewell, Infect. Immun. 64:5117-5128, 1996). Strain CH107 GTF has three internal direct repeats deleted; the 59 carboxyl-terminal amino acids are identical to those of the parental strain. This deletion resulted in decreased enzyme activity but did not affect the amount of cell-associated GTF protein. The GTFs of strains CH2RPE and CH4RPE have six and eight direct repeats, respectively, but are both missing the 14 carboxyl-terminal amino acids. Strain CH2RPE had significantly decreased levels of cell-associated GTF; this decrease was not obviated by the increased number of direct repeats in strain CH4RPE. Thus, the carboxyl-terminal amino acids appeared to influence the amount of cell-associated GTF more than the direct repeats. The qualitative and quantitative differences in the GTFs did not affect the abilities of these strains to accumulate on hydroxyapatite beads in the absence of sucrose. However, when sucrose was added as a substrate for GTF, the mutant strains were unable to accumulate on these surfaces to the same extent as the parent. These differences in sucrose-associated accumulation may be due to changes in the nature of the glucans produced by the different enzymes and/or cohesive interactions between these glucans and the GTF on the surfaces of the growing streptococci.
Vickerman
M M
MM
Department of Microbiology and Immunology, School of Medicine, University of Michigan, Ann Arbor 48109, USA. vickerma@umich.edu
Clewell
D B
DB
D001422:Bacterial Adhesion; D015153:Blotting, Western; D017886:Durapatite / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005964:Glucosyltransferases / Q000032:analysis / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D017384:Sequence Deletion; D013291:Streptococcus / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000276:immunology; D013395:Sucrose / Q000378:metabolism*; D014170:Transformation, Genetic
D013395:Sucrose; D017886:Durapatite; D005964:Glucosyltransferases