Molecular cloning and characterization of a putative mouse hyaluronan synthase
| Autor: | Mary Lou Augustine, Andrew P. Spicer, John A. McDonald |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
DNA
Complementary Protein Conformation Molecular Sequence Data Gene Dosage Molecular cloning Xenopus Proteins Biochemistry Mice Transferases Complementary DNA Animals Amino Acid Sequence RNA Messenger Cloning Molecular Glucuronosyltransferase Hyaluronic Acid Molecular Biology Peptide sequence Gene HAS1 Gene Library biology Base Sequence Sequence Homology Amino Acid cDNA library Glycosyltransferases Membrane Proteins Cell Biology Sequence Analysis DNA Molecular biology Recombinant Proteins Open reading frame Hyaluronan synthase COS Cells biology.protein Hyaluronan Synthases |
| Zdroj: | The Journal of biological chemistry. 271(38) |
| ISSN: | 0021-9258 |
| Popis: | We report the isolation of a novel mouse gene which encodes a putative hyaluronan synthase. The cDNA was identified using degenerate reverse transcriptase-polymerase chain reaction. Degenerate primers were designed based upon an alignment of the amino acid sequences of Streptococcus pyogenes HasA, Xenopus laevis DG42, and Rhizobium meliloti NodC. A mouse embryo cDNA library was screened with the resultant polymerase chain reaction product, and multiple cDNA clones spanning 3 kilobase pairs (kb) were isolated. The open reading frame predicted a 63-kDa protein with several transmembrane sequences, multiple consensus phosphorylation sites, and four putative hyaluronan binding motifs. The amino acid sequence displayed 55% identity to mouse HAS, 56% identity to Xenopus DG42, and 21% identity to Streptococcus HasA. Northern analysis identified transcripts of 4.8 kb and 3.2 kb, which were expressed highly in the developing mouse embryo and at lower levels in adult mouse heart, brain, spleen, lung, and skeletal muscle. Transfection experiments demonstrated that mouse Has2 could direct hyaluronan coat biosynthesis in transfected COS cells, as evidenced by a classical particle exclusion assay. These results suggest that mammalian HA synthase activity is regulated by at least two related genes. Accordingly, we propose the name Has2 for this gene. |
| Databáze: | OpenAIRE |
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