title: Molecular cloning and characterization of an estrogen-dependent porcine oviductal secretory glycoprotein.
abstract: A family of estrogen-dependent porcine oviductal secretory glycoproteins (POSPs) that exhibit structural similarities are synthesized and secreted into the oviductal lumen at proestrus, estrus, and metestrus. The objectives of this study were to clone the POSP cDNA, obtain the full-length cDNA and protein sequence, examine tissue specificity and species distribution, characterize its regulation, and establish its identity by comparison to other known protein, RNA, or DNA sequences. A full-length cDNA of 2022 base pairs was obtained with an open reading frame of 1581 nucleotides, coding for a deduced protein of 527 amino acids (57 970 M(r)). The deduced protein contained three potential N-glycosylation sites, a consensus heparin-binding site, and potential O-glycosylation sites. Amino acid analysis of POSP-E3 confirmed the presence of a 21-amino acid signal sequence. Northern blot analysis revealed an oviduct-specific mRNA species of 2.25 kb in the infundibulum (INF), ampulla (A), and isthmus (I). An mRNA of similar size was detected in the oviduct of the sheep, cow, and rabbit, and one of slightly greater size (2.8 kb) in the mouse and hamster oviduct but not in the horse or alligator oviduct. Dot blot analysis indicated that steady-state levels of POSP mRNA were significantly greater (p = 0.0001) in the A than in the INF or I regardless of day of the estrous cycle and were greater on Day 0 (estrus; p = 0.0001) regardless of location. Further, steady-state mRNA levels were significantly increased (p = 0.02) on Days 0 and 1, declining rapidly to Day 2 through Day 15 of the estrous cycle. Steady-state POSP mRNA levels were significantly greater (p < 0.003) in ovariectomized gilts treated with estradiol valerate than those treated with other steroid regimens, vehicle, or no treatment (Control), consistent with estrogen control of mRNA expression. The POSP protein exhibited significant identity to oviductal glycoproteins from the baboon, cow, hamster, human, mouse, and sheep, to several mammalian nonoviductal glycoproteins; and to several chitinases. POSP joins a growing subfamily of the chitinase gene family that lacks chitinase enzymatic activity.
authors.affiliation: Department of Obstetrics and Gynecology, University of Florida, Gainesvlle 32610, USA. wbuhi.obgyn@obgyn.ufl.edu
authors.forename: W C
authors.initials: WC
authors.lastname: Buhi
authors.forename: I M
authors.initials: IM
authors.lastname: Alvarez
authors.forename: I
authors.initials: I
authors.lastname: Choi
authors.forename: B D
authors.initials: BD
authors.lastname: Cleaver
authors.forename: F A
authors.initials: FA
authors.lastname: Simmen
mesh_terms: D000489:Alligators and Crocodiles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000737:chemistry; D004967:Estrogens / Q000494:pharmacology*; D005260:Female; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D010052:Ovariectomy; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D017385:Sequence Homology; D012756:Sheep; D013045:Species Specificity; D013552:Swine; D014018:Tissue Distribution
chemical_list: D018076:DNA, Complementary; D004967:Estrogens; D006023:Glycoproteins; D012333:RNA, Messenger