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PMID:8366070
| Citation |
DeAngelis, PL, Papaconstantinou, J and Weigel, PH (1993) Molecular cloning, identification, and sequence of the hyaluronan synthase gene from group A Streptococcus pyogenes. J. Biol. Chem. 268:19181-4 |
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| Abstract |
The hyaluronan (HA) synthase of Group A Streptococci has been identified by transposon mutagenesis and deletion analysis. The genes for the HA synthase and a recently identified UDP-Glc dehydrogenase (Dougherty, B. A., and van de Rijn, I. (1993) J. Biol. Chem. 268, 7118-7124) reside on a contiguous stretch of 3.2-kilobase pair DNA that can direct HA biosynthesis in Enterococcus faecalis and Escherichia coli as well as mutant Streptococcus (DeAngelis, P. L., Papaconstantinou, J., and Weigel, P. H. (1993) J. Biol. Chem. 268, 14568-14571). The synthase contains 395 residues (calculated Mr = 45,063) and migrates on SDS-PAGE with a molecular mass of 42 kDa. E. coli K5, which synthesizes UDP-glucuronic acid for production of its endogenous capsular polysaccharide, can make HA if it contains a plasmid encoding the intact 42-kDa protein. E. coli SURE or chi 1448 cells containing the same construct, however, cannot produce HA since these strains cannot make both required sugar nucleotide precursors. The HA synthase is predicted to be an integral membrane protein with four membrane-associated helices, which is consistent with the location of the enzyme activity in Streptococci. There is significant homology between the HA synthase and the Rhizobium nodC gene product, an enzyme that synthesizes chitin-like oligomers. This is the first description at the molecular level of an enzyme shown to synthesize a glycosaminoglycan. |
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| Keywords |
Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA, Fungal/genetics; DNA, Fungal/isolation & purification; Electrophoresis, Polyacrylamide Gel; Enterococcus faecalis/genetics; Escherichia coli/genetics; Genes, Fungal; Glucuronosyltransferase/biosynthesis; Glucuronosyltransferase/genetics; Glucuronosyltransferase/isolation & purification; Glycosyltransferases; Membrane Proteins; Molecular Sequence Data; Molecular Weight; Mutagenesis, Insertional; Oligodeoxyribonucleotides; Plasmids; Recombinant Proteins/biosynthesis; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism; Restriction Mapping; Sequence Homology, Amino Acid; Streptococcus pyogenes/enzymology; Streptococcus pyogenes/genetics; Transferases; Xenopus Proteins |
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Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0050501 : hyaluronan synthase activity |
ECO:0000315: |
F |
Based on fig. 1, hyaluronan(HA) synthase gene was identified based on the fact that Tn insertional mutagenesis caused the band on the gel. Fig. 4 shows the presence of HasA in E. coli with pPD41-delta5 (lane C) and pPD41-deltaPstI(lane D) while table 1 shows positive HA production for strains that contain the plasmid. Strains with pPD41-deltaPstI show little activity for HA synthesis because no HasB present which is required for the reaction. |
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References
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