Abstract

Our goal is to elucidate the structure and molecular mechanisms controlling the expression of terminal deoxynucleotidyl transferase (TdT) during hemopoietic cell differentiation and following neoplastic transformation. TdT is a DNA polymerase that catalyzes polymerization of deoxynucleoside triphosphates without template instruction. Because expression of enzyme activity is restricted to several precursors of lymphocytes and their leukemic counterparts, TdT has been postulated as one source for the generation of somatic mutations and implicated in the deletion of autoreactive lymphocytes. Our overall goal has three principal objectives: (1) elucidation of the structure of the TdT gene in nonexpressing, expressing, and neoplastic cells; (2) determination of mechanisms governing TdT gene expression in normal and neoplastic cells; and (3) determination of the role of posttranslational cleavage and phosphorylation on TdT subcellular localization and enzymatic activity. In addition, an extension of these studies to nonmammalian species will be carried out in order to shed light on the putative correlation between the appearance of the immune system and evolution of the TdT gene. The tools required to pursue these objectives are now available to us. During the current grant period, we constructed and isolated a battery of TdT cDNA probes and monoclonal antibodies upon which our experimental approach is now based. (M)

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA019492-12
Application #
3165160
Study Section
Molecular Biology Study Section (MBY)
Project Start
1976-06-30
Project End
1990-04-30
Budget Start
1987-05-01
Budget End
1988-04-30
Support Year
12
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
University of Kentucky
Department
Type
Schools of Medicine
DUNS #
832127323
City
Lexington
State
KY
Country
United States
Zip Code
40506

  Publications

Yang, B; Gathy, K N; Coleman, M S (1995) T-cell specific avian TdT: characterization of the cDNA and recombinant enzyme. Nucleic Acids Res 23:2041-8
Bhaumik, D; Yang, B; Trangas, T et al. (1994) Identification of a tripartite basal promoter which regulates human terminal deoxynucleotidyl transferase gene expression. J Biol Chem 269:15861-7
Sorscher, D H; Yang, B; Bhaumik, D et al. (1994) Initiation of transcription at the human terminal deoxynucleotidyl transferase gene promoter: a novel role for the TATA binding protein. Biochemistry 33:11025-32
Coleman, M S; Yang, B; Sorscher, D (1992) Regulation of terminal deoxynucleotidyl transferase gene expression in mice and men. Crit Rev Eukaryot Gene Expr 2:237-50
Farrar, Y J; Evans, R K; Beach, C M et al. (1991) Interactions of photoactive DNAs with terminal deoxynucleotidyl transferase: identification of peptides in the DNA binding domain. Biochemistry 30:3075-82
Medin, J A; Hunt, L; Gathy, K et al. (1990) Efficient, low-cost protein factories: expression of human adenosine deaminase in baculovirus-infected insect larvae. Proc Natl Acad Sci U S A 87:2760-4
Evans, R K; Beach, C M; Coleman, M S (1989) Photoaffinity labeling of terminal deoxynucleotidyl transferase. 2. Identification of peptides in the nucleotide binding domain. Biochemistry 28:713-20
Trangas, T; Coleman, M S (1989) Tissue-specific expression of human terminal deoxynucleotidyl transferase is regulated at the transcriptional level. Biochem Biophys Res Commun 164:750-7
Riley, L K; Morrow, J K; Danton, M J et al. (1988) Human terminal deoxyribonucleotidyltransferase: molecular cloning and structural analysis of the gene and 5' flanking region. Proc Natl Acad Sci U S A 85:2489-93
Robbins, D J; Coleman, M S (1988) Initiator role of double stranded DNA in terminal transferase catalyzed polymerization reactions. Nucleic Acids Res 16:2943-57

Showing the most recent 10 out of 13 publications