Expression cloning and molecular characterization of HAS protein, a eukaryotic hyaluronan synthase.

Abstract

We developed a mammalian transient expression system to isolate cDNA clones that determine hyaluronan expression. HAS-, a mouse mammary carcinoma mutant cell line, which is defective in hyaluronan synthase activity, was first established and used as a recipient for the expression cloning. One cloned cDNA that overcame the deficiency was isolated. The cDNA termed HAS contains an open reading frame of 1749 base pairs encoding a new protein of 583 amino acids. Homology analysis of the amino acid sequence suggests that HAS protein is related to streptococcal hyaluronan synthase and also to Xenopus laevis DG42 protein that was found to be homologous to bacterial hyaluronan synthase. Expression of HAS cDNA in HAS- cells complemented not only their mutant phenotypes such as deficient hyaluronan-matrix deposition but also hyaluronan synthase activity itself. Therefore, HAS cDNA is responsible for the activity of the hyaluronan synthase, a key enzyme of hyaluronan synthesis in eukaryotic cells.

• Authors: N Itano, K Kimata
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 271 Issue 17 Pg. 9875-8 (Apr 26 1996) ISSN: 0021-9258 [Print] United States
• PMID: 8626618 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA, Complementary
• Membrane Proteins
• Xenopus Proteins
• Transferases
• Glycosyltransferases
• Glucuronosyltransferase
• HAS1 protein, Xenopus
• Hyaluronan Synthases

Topics

• Amino Acid Sequence
• Animals
• Base Sequence
• Cloning, Molecular
• DNA, Complementary (genetics)
• Glucuronosyltransferase (chemistry, genetics)
• Glycosyltransferases
• Hyaluronan Synthases
• Membrane Proteins
• Mice
• Molecular Sequence Data
• Sequence Alignment
• Sequence Homology, Amino Acid
• Solubility
• Transfection
• Transferases
• Xenopus Proteins