Abstract

This is a once-amended proposal to continue Dr. Smulson's studies into Three aims are proposed to pursue this objective. The first is structured to attempt to clarify the interaction of PADPRP with DNA breaks in chromatin. Initial studies will evaluate the cycling of PADPRP on and off ends of DNA during repair, using bacterially-expressed PADPRP mutant proteins. Also, using stably transfected synchronized 3T3-L1 cells, evidence will be sought to establish the direct role of PADPRP with DNA replication/repair by defining the physical association with DNA polymerase alpha, as well as a multi-enzyme DNA replicative complex. The cleavage of PADPRP during apoptosis in this system also will be evaluated.
Aim II is designed to determine whether the poly(ADP-ribose) signal generated subsequent to DNA strand breaks plays a significant role in the accumulation of p53 protein and the arrest of cells at G1 until repair can proceed.
In aim III the mechanistic role of PADPRP in V(D)J recombination, which shares several enzymatic features with strand break rejoining during repair, will be explored. The effects of the PADPRP expression constructs on this process will be examined in mouse 3T3 cells expressing the RAG 1 and RAG 2 genes and in the context of the whole animal by use of newly-established """"""""knockout"""""""" mice.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA025344-19
Application #
2653960
Study Section
Chemical Pathology Study Section (CPA)
Program Officer
Okano, Paul
Project Start
1979-06-01
Project End
2000-01-31
Budget Start
1998-02-01
Budget End
1999-01-31
Support Year
19
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
Georgetown University
Department
Biochemistry
Type
Schools of Dentistry
DUNS #
049515844
City
Washington
State
DC
Country
United States
Zip Code
20057

  Publications

Wang, Tao; Simbulan-Rosenthal, Cynthia M; Smulson, Mark E et al. (2008) Polyubiquitylation of PARP-1 through ubiquitin K48 is modulated by activated DNA, NAD+, and dipeptides. J Cell Biochem 104:318-28
Rosenthal, D S; Simbulan-Rosenthal, C M; Liu, W F et al. (2001) Mechanisms of JP-8 jet fuel cell toxicity. II. Induction of necrosis in skin fibroblasts and keratinocytes and modulation of levels of Bcl-2 family members. Toxicol Appl Pharmacol 171:107-16
Stoica, B A; Boulares, A H; Rosenthal, D S et al. (2001) Mechanisms of JP-8 jet fuel toxicity. I. Induction of apoptosis in rat lung epithelial cells. Toxicol Appl Pharmacol 171:94-106
Smulson, M; Istock, N; Ding, R et al. (1994) Deletion mutants of poly(ADP-ribose) polymerase support a model of cyclic association and dissociation of enzyme from DNA ends during DNA repair. Biochemistry 33:6186-91
Stevnsner, T; Ding, R; Smulson, M et al. (1994) Inhibition of gene-specific repair of alkylation damage in cells depleted of poly(ADP-ribose) polymerase. Nucleic Acids Res 22:4620-4
Ding, R; Smulson, M (1994) Depletion of nuclear poly(ADP-ribose) polymerase by antisense RNA expression: influences on genomic stability, chromatin organization, and carcinogen cytotoxicity. Cancer Res 54:4627-34
Smulson, M E (1994) Poly(ADP-ribose) polymerase gene on chromosome 1q: early role in differentiation linked replication;gene on human chromosome 13q: marker of carcinogenesis. Mol Cell Biochem 138:77-84
Blaho, J A; Michael, N; Kang, V et al. (1992) Differences in the poly(ADP-ribosyl)ation patterns of ICP4, the herpes simplex virus major regulatory protein, in infected cells and in isolated nuclei. J Virol 66:6398-407
Ding, R; Pommier, Y; Kang, V H et al. (1992) Depletion of poly(ADP-ribose) polymerase by antisense RNA expression results in a delay in DNA strand break rejoining. J Biol Chem 267:12804-12
Bhatia, K; Kang, V H; Stein, G S et al. (1990) Cell cycle regulation of an exogenous human poly(ADP-ribose) polymerase cDNA introduced into murine cells. J Cell Physiol 144:345-53

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