Evidence has been obtained that an estrogen=induced chick oviduct membrane glycoprotein is the transferrin receptor. Estrogen treatment elicits oviduct differentiation which includes the biosynthesis of egg white proteins. One of the egg white proteins, conalbumin (ovotransferrin), serves as an iron source for the developing embryo. In order to obtain the large amounts of iron necessary for conalbumin secretion the oviduct transferrin receptor is also estrogen-induced. Antibodies and/or a specific oligonucleotide probe (based on N-terminal sequence analysis) will be employed to obtain a transferrin receptor cDNA from a lambda gtll expression library. The cDNA will be used to obtain protein sequence and to measure steady-state mRNA levels, transcriptional activity of receptor mRNA and mRNA half-life. The segregation of transferrin receptor from conalbumin (ovotransferrin) during movement from their-site(s) of synthesis to the cell surface will be studied using transferrin receptor antibodies, conalbumin antibodies and immunoelectron microscopy with gold- or ferritin- conjugated secondary antibodies. Additionally, an attempt will be made to obtain the putative transferrin receptor-specific vesicles by sucrose gradient centrifugation. Depending on the outcome of the segregation studies additional experiments may be undertaken to determine when the transferrin receptor acquires its binding capability and how that might affect vesicular segregation.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
2R01HD009046-12A2
Application #
3311043
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1978-06-01
Project End
1993-03-31
Budget Start
1988-07-01
Budget End
1989-03-31
Support Year
12
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Upstate Medical University
Department
Type
Schools of Medicine
DUNS #
058889106
City
Syracuse
State
NY
Country
United States
Zip Code
13210
Cho, S S; Lucas, J J (1995) Immunocytochemical study with an anti-transferrin binding protein serum: a marker for avian oligodendrocytes. Brain Res 674:15-25
Hayes, G R; Himpler, B S; Weiner, K X et al. (1994) A chicken transferrin binding protein is heat shock protein 108. Biochem Biophys Res Commun 200:65-70
Fuernkranz, H A; Schwob, J E; Lucas, J J (1991) Differential tissue localization of oviduct and erythroid transferrin receptors. Proc Natl Acad Sci U S A 88:7505-8
Poola, I; Mason, A B; Lucas, J J (1990) The chicken oviduct and embryonic red blood cell transferrin receptors are distinct molecules. Biochem Biophys Res Commun 171:26-32
Seifert, S C; Lucas, J J (1988) Incorporation of mevalonate into dolichol and other isoprenoids during estrogen-induced chick oviduct differentiation. Biochim Biophys Acta 962:16-24
Poola, I; Lucas, J J (1988) Purification and characterization of an estrogen-inducible membrane glycoprotein. Evidence that it is a transferrin receptor. J Biol Chem 263:19137-46
Lucas, J J; Makunike, C; Basinger, S F et al. (1986) Localization of the lipid intermediate pathway of protein glycosylation in oviduct cell types. Tissue Cell 18:241-9
Starr, C M; Lucas, J J (1985) Regulation of dolichyl phosphate-mediated protein glycosylation: estrogen effects on glucosyl transfers in oviduct membranes. Arch Biochem Biophys 237:261-70
Malvar, T M; Hallahan, T W; Beach, D H et al. (1985) Hydrolysis of dolichyl esters by oviduct membranes and characterization of endogenous dolichyl esters. Arch Biochem Biophys 238:401-9