Pregnancy specific Beta 1 glycoprotein (SP1) besides being a good index for monitoring abnormal pregnancies, fetal well being and as a marker for trophoblastic tumors, may also play a very important role in implantation of the embryo and the subsequent growth of the fetus. The long-term objective of this proposal is to understand the functional roles of SP1 in human pregnancy with the view to enhance our knowledge of factors which effect the well-being of the growing fetus. The major obstacle to achieve this goal in the past is the occurrence of molecular heterogeneity of the SP1 protein making all the immunochemical assaying methods for its quantitation unreliable. This proposal specifically will (1) Try to solve the question of molecular heterogeneity of SP1 by cloning the cDNA of the protein. SP1 cDNA clones will be obtained by screening a human placental cDNA expression library using labelled antibody. The identity of the clones obtained will be confirmed by techniques including Western blot hybridization, genomic blot hybridization and DNA and protein sequencing; (2) Investigate the validity of cultured human skin fibroblasts, cultured placental cells, mouse, rat and baboon serving as models for studying SP1. The SP1 like molecules in these systsems will be compared with human SP1 by Northern blot hybridization first. Further comparison will be made by cloning the cDNA of these molecules and compared with human SP1 cDNA by restriction enzyme mapping and, when needed, by DNA sequencing; (3) Investigate the mechanism of enhanced SP1 production in Meckel's syndrome. Cultured fibroblasts derived from a patient will be used. The production of SP1 in these fibroblasts will be studied by immunoprecipitation. If expression of the genetic defect in the fibroblast is obvious, mechanism of SP1 overproduction will be further investigated by quantitating the mRNA of SP1 with dot blot hybridization. The cDNA will then be cloned and examined by restriction enzyme mapping and eventually DNA sequencing; (4) Determine the gene structure of SP1. This will be achieved by screening a human genomic library with SP1 cDNA probe. The clones hybridizing to the probe will be picked and sequenced. The gene structure of SP1 will be compared with that of human placental lactogen and human chorionic gonadotropin Beta chain. With techniques such as chromosomal walking, genomic blot hybridizing etc., the arrangement of these genes on the human genome will also be determined; (5) Lastly, investigate the properties of SP1 like molecules in tumor tissues using hydatidiform mole as an example. Techniques used will include Northern blot hybridization to identify the presence of SP1, dot blot hybridization to quantitate the quantity of mRNA produced, cloning restriction enzyme map and DNA sequencing to determine the structure and nature of the SP1 protein in tumor.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
7R01HD021793-02
Application #
3320923
Study Section
Human Embryology and Development Subcommittee 2 (HED)
Project Start
1987-04-01
Project End
1989-03-31
Budget Start
1987-04-01
Budget End
1988-03-31
Support Year
2
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Oklahoma Medical Research Foundation
Department
Type
DUNS #
937727907
City
Oklahoma City
State
OK
Country
United States
Zip Code
73104
Blomberg, L A; Cohn, M L; Cahill, R A et al. (1998) Effect of human pregnancy-specific beta1-glycoprotein on blood cell regeneration after bone marrow transplantation. Proc Soc Exp Biol Med 217:212-8
Shupert, W L; Chan, W Y (1993) Pregnancy specific beta 1-glycoprotein in human intestine. Mol Cell Biochem 120:159-70
Ida, H; Rennert, O M; Eto, Y et al. (1993) Cloning of a human acid sphingomyelinase cDNA with a new mutation that renders the enzyme inactive. J Biochem 114:15-20
Wu, S M; Bazar, L S; Cohn, M L et al. (1993) Expression of pregnancy-specific beta 1-glycoprotein genes in hematopoietic cells. Mol Cell Biochem 122:147-58
Chan, W Y; Zheng, Q X; McMahon, J et al. (1991) Characterization of new members of the pregnancy-specific beta 1-glycoprotein family. Mol Cell Biochem 106:161-70
Borjigin, J; Tease, L A; Barnes, W et al. (1990) Expression of the pregnancy-specific beta 1-glycoprotein genes in human testis. Biochem Biophys Res Commun 166:622-9
Zheng, Q X; Tease, L A; Shupert, W L et al. (1990) Characterization of cDNAs of the human pregnancy-specific beta 1-glycoprotein family, a new subfamily of the immunoglobulin gene superfamily. Biochemistry 29:2845-52
Ogilvie, S; Shiverick, K T; Larkin, L H et al. (1990) Pregnancy-specific beta 1-glycoprotein messenger ribonucleic acid and immunoreactive protein in the rat testis. Endocrinology 126:292-8
Tease, L A; Fazleabas, A T; Chan, W Y (1989) Characterization of baboon pregnancy-specific beta 1-glycoprotein. Biol Reprod 41:1113-21
Chan, W Y; Liu, Q R; Borjigin, J et al. (1989) Characterization of the cDNA encoding human nucleophosmin and studies of its role in normal and abnormal growth. Biochemistry 28:1033-9

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