Changes in the carboxyl-terminal repeat region affect extracellular activity and glucan products of Streptococcus gordonii glucosyltransferase. 1996

M M Vickerman, and M C Sulavik, and P E Minick, and D B Clewell
Department of Microbiology and Immunology, School of Medicine, University of Michigan, Ann Arbor 48109, USA. vickerma@umich.edu

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.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D012091 Repetitive Sequences, Nucleic Acid Sequences of DNA or RNA that occur in multiple copies. There are several types: INTERSPERSED REPETITIVE SEQUENCES are copies of transposable elements (DNA TRANSPOSABLE ELEMENTS or RETROELEMENTS) dispersed throughout the genome. TERMINAL REPEAT SEQUENCES flank both ends of another sequence, for example, the long terminal repeats (LTRs) on RETROVIRUSES. Variations may be direct repeats, those occurring in the same direction, or inverted repeats, those opposite to each other in direction. TANDEM REPEAT SEQUENCES are copies which lie adjacent to each other, direct or inverted (INVERTED REPEAT SEQUENCES). DNA Repetitious Region,Direct Repeat,Genes, Selfish,Nucleic Acid Repetitive Sequences,Repetitive Region,Selfish DNA,Selfish Genes,DNA, Selfish,Repetitious Region, DNA,Repetitive Sequence,DNA Repetitious Regions,DNAs, Selfish,Direct Repeats,Gene, Selfish,Repeat, Direct,Repeats, Direct,Repetitious Regions, DNA,Repetitive Regions,Repetitive Sequences,Selfish DNAs,Selfish Gene
D004269 DNA, Bacterial Deoxyribonucleic acid that makes up the genetic material of bacteria. Bacterial DNA
D005936 Glucans Polysaccharides composed of repeating glucose units. They can consist of branched or unbranched chains in any linkages. Glucan,Polyglucose,Polyglucoses,Glucan (BO),Glucose Polymer,Polycose,Polymer, Glucose
D005964 Glucosyltransferases Enzymes that catalyze the transfer of glucose from a nucleoside diphosphate glucose to an acceptor molecule which is frequently another carbohydrate. EC 2.4.1.-. Glucosyltransferase
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D013291 Streptococcus A genus of gram-positive, coccoid bacteria whose organisms occur in pairs or chains. No endospores are produced. Many species exist as commensals or parasites on man or animals with some being highly pathogenic. A few species are saprophytes and occur in the natural environment.

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