The aim of the proposed research is to study the chemotherapy of cancer at the level of gene transcription. We have previously shown that DNA-dependent RNA polymerase II in chicken myeloblastosis leukemic cells is different both quantitatively and qualitatively from that in normal chicken bone marrow cells. We have also isolated a protein factor from the chicken leukemic cells, whch stimulates the initiation of in vitro RNA synthesis by RNA polymerase II. We propose to further purify the RNA polymerase and its initiation factor to homogeneity. The purified enzymes from leukemic as well as normal cells will be studied with regard to their properties, functions, subunit structures and reconstruction, and analysis of their product RNA. This is expected to provide further evidence for a leukemic-specific RNA polymerase. The purified RNA polymerase initiation factor will be studied to determine its properties, physiological functions and the mechanism by which it stimulates the initiation of RNA synthesis. Attempts will also be made to determine its activity in other normal and malignant cells (normal chicken meloblast cells, avian myeloblastosis viral non-producer cell line, human leukemic and human blyphoma cell lines). These studies will reveal whether the factor is specific for the gene expression of cancerous growth or it is universally required for the RNA synthesis of eukaryotic cells. We propose to continue our studies on several currently used and newer experimental antineoplastic agents for thier inhibitory effects on neoplastic gene transcription, as well as the biochemical mechanisms of the inhibition. The agents used in this proposal include cytosine arabinoside, notrosourea compounds, adriamaycin and its derivatives (AD 32, AD 41 and AD 142), 6-mercaptopurine and its methylated derivatives. The ultimate goal of the proposed research is to determine if it is possible to arrest neoplastic expression of cancer cells at transcriptional level by selectively inhibiting the activity of malignant RNA polymerase (or its regulatory factor) without affecting the normal enzyme.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA033022-04
Application #
3170945
Study Section
Experimental Therapeutics Subcommittee 2 (ET)
Project Start
1982-07-01
Project End
1988-07-31
Budget Start
1985-09-01
Budget End
1986-07-31
Support Year
4
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of California Davis
Department
Type
Schools of Medicine
DUNS #
094878337
City
Davis
State
CA
Country
United States
Zip Code
95618
Chuang, L F; Zhao, F K; Chuang, R Y (1989) Isolation and purification of protein kinase C from human leukemia ML-1 cells phosphorylation of human leukemia RNA polymerase II in vitro. Biochim Biophys Acta 992:87-95
Zhao, F K; Chuang, L F; Israel, M et al. (1989) Adriamycin interacts with diacylglycerol to inhibit human leukemia protein kinase C. Anticancer Res 9:225-9
Zhao, F K; Chuang, L F; Israel, M et al. (1989) Cremophor EL, a widely used parenteral vehicle, is a potent inhibitor of protein kinase C. Biochem Biophys Res Commun 159:1359-67
Wang, L F; Doi, R H; Chuang, L F et al. (1989) Bluetongue virus-17 fusion protein ns1 expressed in Escherichia coli by pUC vectors. Biochem Biophys Res Commun 162:892-9
Lameh, J; Chuang, R Y; Israel, M et al. (1989) Nucleoside uptake and membrane fluidity studies on N-trifluoroacetyladriamycin-14-O-hemiadipate-treated human leukemia and lymphoma cells. Cancer Res 49:2905-8
Chuang, L F; Chuang, R Y (1988) The 24,000 Da subunit is not required for the RNA synthesis activity of chicken leukemia RNA polymerase II. Biochimie 70:1397-400
Lameh, J; Chuang, L F; Israel, M et al. (1988) Mechanistic studies on N-benzyladriamycin-14-valerate (AD 198), a highly lipophilic alkyl adriamycin analog. Anticancer Res 8:689-93
Wu, J C; Chernow, M; Boehme, R E et al. (1988) Kinetics and inhibition of reverse transcriptase from human and simian immunodeficiency viruses. Antimicrob Agents Chemother 32:1887-90
Chuang, L F; Chuang, R Y; Acton, E M et al. (1987) Effect of morpholinyladriamycin analogs and adriamycin on the activities of DNA polymerase alpha and RNA polymerase II of chicken leukemia cells. J Pharmacol Exp Ther 242:372-7
Chuang, L F; Cooper, R H; Yau, P et al. (1987) Protein kinase C phosphorylates leukemia RNA polymerase II. Biochem Biophys Res Commun 145:1376-83

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