We are investigating the molecular mechanisms of p53-mediated apoptosis. Our previous studies have identified a novel pathway of apoptosis involving the functional and physical interaction of p53 with DNA helicases, XPB and XPD. We have extended these studies to other members of the RecQ helicase family, BLM and WRN, that are linked to cancer predisposition syndromes, Bloom and Werner, respectively. Germline mutations in WRN are found in patients with the premature aging and cancer susceptibility syndrome known as Werner syndrome (WS). p53 binds to the WRN protein in vivo and in vitro through its carboxyl terminus. WS fibroblasts have an attenuated p53-mediated apoptotic response, and this deficiency can be rescued by expression of wild-type WRN. These data support the hypothesis that p53 can induce apoptosis through the modulation of specific DExH-containing DNA helicases and may have implications for the cancer predisposition observed in WS patients.Bloom syndrome (BS) is an autosomal recessive genomic instability syndrome characterized by growth retardation, immune deficiency and cancer predisposition. Similar to cells from XPB, XPD or WS individuals who have an attenuated p53-dependent apoptotic pathway, p53-mediated apoptosis also is defective in BS fibroblasts. This apoptotic pathway can be functionally rescued by the expression of the wild-type (wt) BLM gene. Lymphoblastoid cell lines (LCLs) derived from BS donors are resistant to either gamma-radiation or adriamycin-induced cell killing, and also can be rescued by the wt BLM. In contrast, BS cells have a normal Fas-mediated apoptosis, and a normal DNA damage- induced p53 accumulation and G1-S and G2-M cell cycle checkpoints. BLM localizes in nuclear foci identified as PML nuclear bodies (NBs), a structure that also contains the promyelocytic leukemia protein (PML), Rb, SUMO-1 and others, and may be involved in apoptosis. Cells from Li- Fraumeni syndrome (LFS) patients carrying p53 germline mutations have a decreased number of BLM foci. The induction of p53 increased the number of BLM foci, but did not alter either BLM levels or the number of NBs. These results indicate a novel function of p53 and are consistent with the hypothesis that, nuclear trafficking of BLM to NBs mediated by p53, contributes to its apoptotic activity. - Apoptosis, DNA repair, p, , Tumor Suppressor, - Human Tissues, Fluids, Cells, etc.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Intramural Research (Z01)
Project #
1Z01BC005794-05
Application #
6289169
Study Section
Special Emphasis Panel (LHC)
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
1999
Total Cost
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Hussain, S P; Raja, K; Amstad, P A et al. (2000) Increased p53 mutation load in nontumorous human liver of wilson disease and hemochromatosis: oxyradical overload diseases. Proc Natl Acad Sci U S A 97:12770-5
Hsia, C C; Nakashima, Y; Thorgeirsson, S S et al. (2000) Correlation of immunohistochemical staining and mutations of p53 in human hepatocellular carcinoma. Oncol Rep 7:353-6
Hussain, S P; Amstad, P; Raja, K et al. (2000) Increased p53 mutation load in noncancerous colon tissue from ulcerative colitis: a cancer-prone chronic inflammatory disease. Cancer Res 60:3333-7
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Bennett, W P; Hussain, S P; Vahakangas, K H et al. (1999) Molecular epidemiology of human cancer risk: gene-environment interactions and p53 mutation spectrum in human lung cancer. J Pathol 187:8-18
Hussain, S P; Harris, C C (1999) p53 mutation spectrum and load: the generation of hypotheses linking the exposure of endogenous or exogenous carcinogens to human cancer. Mutat Res 428:23-32
Hagiwara, K; McMenamin, M G; Miura, K et al. (1999) Mutational analysis of the p63/p73L/p51/p40/CUSP/KET gene in human cancer cell lines using intronic primers. Cancer Res 59:4165-9
Lebel, M; Spillare, E A; Harris, C C et al. (1999) The Werner syndrome gene product co-purifies with the DNA replication complex and interacts with PCNA and topoisomerase I. J Biol Chem 274:37795-9
Spillare, E A; Robles, A I; Wang, X W et al. (1999) p53-mediated apoptosis is attenuated in Werner syndrome cells. Genes Dev 13:1355-60
Robles, A I; Wang, X W; Harris, C C (1999) Drug-induced apoptosis is delayed and reduced in XPD lymphoblastoid cell lines: possible role of TFIIH in p53-mediated apoptotic cell death. Oncogene 18:4681-8

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