Molecular cloning and temporal expression during pregnancy of the messenger ribonucleic acid encoding uteroferrin, a progesterone-induced uterine secretory protein.
A lambda gt11 expression library containing cDNA inserts prepared from porcine endometrial mRNA was immunologically screened by using an antiserum developed against porcine uteroferrin (Uf), a glycoprotein that has been strongly implicated in transplacental iron transport in the pregnant pig. Antibody reactive clones (lambda 4a3, 13.1, and 2.2) were isolated after screening 1.5 x 10(5) recombinant phages. Clones 4a3 and 13.1 expressed Uf antigenic determinants in beta-galactosidase fusion proteins and specifically selected antibody which reacted with Uf in immunoblots prepared from uterine cytosolic extracts. In addition, all three cDNA clones collectively contained DNA sequences that encoded an 85-amino acid peptide which corresponded to a region within the carboxyterminal portion of the Uf protein. Northern blot hybridization of these cDNAs to RNAs extracted from whole uterine tissue of pregnant pigs revealed a single uterine poly(A)+ RNA of approximately 1.7 kilobases in length, which was not found in liver and mammary tissue RNAs. The concentration of the Uf mRNA changed in a temporal fashion during pregnancy in a manner that was distinct from that of the progesterone receptor mRNAs. Highest levels of Uf mRNA were found at mid and late pregnancy (days 45-110) and were about 50-fold greater than at day 30 of pregnancy. By contrast, RIA analysis of the uterine tissue extracts showed that maximum amounts of Uf were present at day 60 and then declined sharply. Thus the pattern of Uf mRNA present in the uterus did not parallel the amount of Uf polypeptide that could be recovered from the tissue. The tissue specific and temporal regulation of Uf gene expression emphasizes that the protein plays an important role in uterine activity and/or fetal development.
Simmen
R C
RC
Department of Animal Science, Ohio State University, Wooster 44691.
Baumbach
G A
GA
Roberts
R M
RM
D000135:Acid Phosphatase; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D005260:Female; D005786:Gene Expression Regulation; D007120:Immunochemistry; D007527:Isoenzymes; D008667:Metalloproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D011247:Pregnancy; D011270:Pregnancy, Animal; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics; D013552:Swine; D000071681:Tartrate-Resistant Acid Phosphatase; D014599:Uterus / Q000378:metabolism
D000906:Antibodies; D007527:Isoenzymes; D008667:Metalloproteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase