title: Molecular cloning, characterization and isolation of novel spliced variants of the human ortholog of a rat estrogen-regulated membrane-associated protein, UO-44.
abstract: 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 HuUO-44 transcripts of approximately 2 and 3 kb in the pancreas. RT-PCR demonstrated that HuUO-44 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 HuUO-44 are two CUB domains and a zona-pellucida domain. Transfection studies demonstrated the membrane-associated nature of HuUO-44. By immunohistochemistry, HuUO-44 was located to the normal ovarian and ovarian tumor epithelial cells; in NIH-OVCAR3 ovarian cancer cells, HuUO-44 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 HuUO-44 antiserum. These findings suggest the potential role of HuUO-44 in cell motility, cell-cell interactions and/or interactions with the extracellular matrices.
authors.affiliation: Laboratory of Molecular Endocrinology, Division of Cellular and Molecular Research, National Cancer Centre of Singapore, Singapore 169610, Singapore.
authors.forename: Caine Tuck Choy
authors.initials: CT
authors.lastname: Leong
authors.forename: Chuan Young
authors.initials: CY
authors.lastname: Ng
authors.forename: Choon Kiat
authors.initials: CK
authors.lastname: Ong
authors.forename: Chee Pang
authors.initials: CP
authors.lastname: Ng
authors.forename: Zeng Shuan
authors.initials: ZS
authors.lastname: Ma
authors.forename: Thanh Hung
authors.initials: TH
authors.lastname: Nguyen
authors.forename: Sun Kuie
authors.initials: SK
authors.lastname: Tay
authors.forename: Hung
authors.initials: H
authors.lastname: Huynh
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D001483:Base Sequence; D002450:Cell Communication; D002455:Cell Division; D002465:Cell Movement; D002879:Chromosomes, Human, Pair 10; D003001:Cloning, Molecular; D004967:Estrogens / Q000494:pharmacology*; D005260:Female; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010051:Ovarian Neoplasms / Q000378:metabolism; D010053:Ovary / Q000378:metabolism; D020033:Protein Isoforms; D014162:Transfection
chemical_list: C488236:CUZD1 protein, human; D004967:Estrogens; D008565:Membrane Proteins; D020033:Protein Isoforms