Abstract

Studies are planned to continue to investigate the structure, expression, and function of tropomyosin in normal and transformed cells. Full-length cDNA clones encoding all five isoforms of tropomyosin from rat embryonic fibroblasts will be isolated and their DNA sequences determined to deduce their primary structure. This information will be used to prepare synthetic peptides to be used as immunogens for making isoform-specific antibodies. Such antibodies will be useful in determining the subcellular distribution of each form of tropomyosin and if this localization is altered upon transformation. Bacteriophage lambda recombinants which carry the genes for rat tropomyosin will be isolated and the general structure and organization of the genes determined. For example, these studies will determine if all five forms of fibroblast tropomyosin arise from separate but related genes or if some arise from a single gene via differential processing of the message. The cloned DNAs will be used to study tropomyosin expression to determine if there are alterations in transcription, processing, and translation of tropomyosin messages in transformed cells. To study the functional significance of the altered pattern of tropomyosins in transformed fibroblasts, tropomyosin will be introduced into living cells by (1) microinjection of purified mRNA or (2) the expression of cloned DNAs encoding tropomyosin, for example, via the metallothionein promoter. These studies will determine if expression of tropomyosin in transformed cells will result in changes in cell shape or organization of microfilaments. (V)

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
5P01CA029569-05
Application #
3093300
Study Section
Cancer Special Program Advisory Committee (CAK)
Project Start
1981-04-01
Project End
1986-03-31
Budget Start
1985-04-01
Budget End
1986-03-31
Support Year
5
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Cold Spring Harbor Laboratory
Department
Type
DUNS #
065968786
City
Cold Spring Harbor
State
NY
Country
United States
Zip Code
11724

  Publications

Addison, W R; Kurtz, D T (1989) Identification of nuclear proteins that bind to the glucocorticoid regulatory region of a rat alpha 2u-globulin gene. J Biol Chem 264:21891-5
MacInnes, J I; Nozik, E S; Kurtz, D T (1986) Tissue-specific expression of the rat alpha 2u globulin gene family. Mol Cell Biol 6:3563-7
Helfman, D M; Cheley, S; Kuismanen, E et al. (1986) Nonmuscle and muscle tropomyosin isoforms are expressed from a single gene by alternative RNA splicing and polyadenylation. Mol Cell Biol 6:3582-95
Addison, W R; Kurtz, D T (1986) Nucleotide sequences required for the regulation of a rat alpha 2u-globulin gene by glucocorticoids. Mol Cell Biol 6:2334-46
Krangel, M S (1985) Unusual RNA splicing generates a secreted form of HLA-A2 in a mutagenized B lymphoblastoid cell line. EMBO J 4:1205-10
Gross, M; Sweet, R W; Sathe, G et al. (1985) Purification and characterization of human H-ras proteins expressed in Escherichia coli. Mol Cell Biol 5:1015-24
Yamawaki-Kataoka, Y; Helfman, D M (1985) Rat embryonic fibroblast tropomyosin 1. cDNA and complete primary amino acid sequence. J Biol Chem 260:14440-5
Catelli, M G; Binart, N; Feramisco, J R et al. (1985) Cloning of the chick hsp 90 cDNA in expression vector. Nucleic Acids Res 13:6035-47
Birchmeier, C; Broek, D; Wigler, M (1985) ras proteins can induce meiosis in Xenopus oocytes. Cell 43:615-21
Birkenmeier, C S; Bodine, D M; Repasky, E A et al. (1985) Remarkable homology among the internal repeats of erythroid and nonerythroid spectrin. Proc Natl Acad Sci U S A 82:5671-5