4.8 Article

Molecular cloning, characterization and isolation of novel spliced variants of the human ortholog of a rat estrogen-regulated membrane-associated protein, UO-44

Journal

ONCOGENE
Volume 23, Issue 33, Pages 5707-5718

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/sj.onc.1207754

Keywords

estrogen regulated; membrane associated; CUB domains; zona-pellucida and HuUO-44

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We have previously reported the characterization of an estrogen-regulated rat uterine-ovarian-specific complementary DNA (UO-44). To understand the involvement of this protein in the initiation and progression of human ovarian and uterine cancers, we now report the cloning and characterization of the human ortholog (HuUO-44). HuUO-44 is mapped to chromosome 10q26.13 and contains nine exons. Multiple tissue Northern blot detected two HuU044 transcripts of approximately 2 and 3kb in the pancreas. RT-PCR demonstrated that HuU044 undergoes a complex series of alternative splicing events between exons 2 and 6 that yielded four novel splice variants, HuUO-44A, HuUO-44B, HuUO-44C and HuUO-44D. Putative functional motifs identified in HuU044 are two CUB domains and a zona-pellucida domain. Transfection studies demonstrated the membrane-associated nature of HuU044. By immunohistochemistry, HuU044 was located to the normal ovarian and ovarian tumor epithelial cells; in NIH-OVCAR3 ovarian cancer cells, HuU044 was detected only at the leading edge of the dividing cells. Most importantly, a marked loss in cell attachment and proliferation was observed in NIH-OVCAR3 cells cultured in the presence of a polyclonal HuU044 antiserum. These findings suggest the potential role of HuU044 in cell motility, cell-cell matrices.

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