Regulation of hyaluronidase activity by alternative mRNA splicing

Vinata B Lokeshwar, Grethchen L. Schroeder, Robert I. Carey, Mark S. Soloway, Naoko Iida

Research output: Contribution to journalArticle

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Abstract

Hyaluronidase is a hyaluronic acid-degrading endoglycosidase that is present in many toxins and the levels of which are elevated in cancer. Increased concentration of HYAL1-type hyaluronidase correlates with tumor progression and is a marker for grade (G) 2 or 3 bladder cancer. Using bladder tissues and cells, prostate cancer cells, and kidney tissues and performing reverse transcription-PCR, cDNA cloning, DNA sequencing, and in vitro translation, we identified splice variants of HYAL1 and HYAL3. HYAL1v1 variant lacks a 30-amino acid (aa) sequence (301-330) present in HYAL1 protein. HYAL1v1, HYAL1v2 (aa 183-435 present in HYAL1 wild type), HYAL1v3 (aa 1-207), HYAL1v4 (aa 260-435), and HYAL1v5 (aa 340-435) are enzymatically inactive and are expressed in normal tissues/cells and G1 bladder tumor tissues. However, HYAL1 wild type is expressed in G2/G3 tumors and in invasive tumor cells. Stable transfection and HYAL1v1-specific antibody confirmed that the HYAL1 sequence from aa 301 to 330 is critical for hyaluronidase activity. All tumor cells and tissues mainly express HYAL3 variants. HYAL3v1 lacks a 30-aa sequence (299-328) present in HYAL3 protein, that is homologous to the 30-aa HYAL1 sequence. HYAL3v1, HYAL3v2 (aa 251-417 present in HYAL3 wild type), and HYAL3v3 (aa 251-417, but lacking aa 299-328), are enzymatically inactive. Although splicing of a single independent exon generates HYAL1v1 and HYAL3v1, internal exon splicing generates the other HYAL1/HYAL3 variants. These results demonstrate that alternative mRNA splicing controls cellular expression of enzymat. ically active hyaluronidase and may explain the elevated hyaluronidase levels in bladder/prostate cancer.

Original languageEnglish (US)
Pages (from-to)33654-33663
Number of pages10
JournalJournal of Biological Chemistry
Volume277
Issue number37
DOIs
StatePublished - Sep 13 2002
Externally publishedYes

Fingerprint

Hyaluronoglucosaminidase
Alternative Splicing
Amino Acids
Messenger RNA
Amino Acid Sequence
Urinary Bladder Neoplasms
Tumors
Tissue
Neoplasms
Exons
Prostatic Neoplasms
Cells
Kidney Neoplasms
Glycoside Hydrolases
Hyaluronic Acid
DNA Sequence Analysis
Reverse Transcription
Transfection
Organism Cloning
Urinary Bladder

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Regulation of hyaluronidase activity by alternative mRNA splicing. / Lokeshwar, Vinata B; Schroeder, Grethchen L.; Carey, Robert I.; Soloway, Mark S.; Iida, Naoko.

In: Journal of Biological Chemistry, Vol. 277, No. 37, 13.09.2002, p. 33654-33663.

Research output: Contribution to journalArticle

Lokeshwar, Vinata B ; Schroeder, Grethchen L. ; Carey, Robert I. ; Soloway, Mark S. ; Iida, Naoko. / Regulation of hyaluronidase activity by alternative mRNA splicing. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 37. pp. 33654-33663.
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