Abstract

The p53 tumor suppressor regulates diverse antiproliferative processes such that cells acquiring p53 mutations have impaired cell-cycle checkpoints, senescence, apoptosis, and genomic stability. Here, we use stable RNA interference to examine the role of PUMA, a p53 target gene and proapoptotic member of the Bcl2 family, in p53-mediated tumor suppression. PUMA short hairpin RNAs (shRNAs) efficiently suppressed PUMA expression and p53-dependent apoptosis but did not impair nonapoptotic functions of p53. Like p53 shRNAs, PUMA shRNAs promoted oncogenic transformation of primary murine fibroblasts by the E1A/ras oncogene combination and dramatically accelerated myc-induced lymphomagenesis without disrupting p53-dependent cell-cycle arrest. However, the ability of PUMA to execute p53 tumor suppressor functions was variable because, in contrast to p53 shRNAs, PUMA shRNAs were unable to cooperate with oncogenic ras in transformation. These results demonstrate that the p53 effector functions involved in tumor suppression are context dependent and, in some settings, depend heavily on the expression of a single proapoptotic effector. Additionally, they demonstrate the utility of RNA interference for evaluating putative tumor suppressor genes in vivo.

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Acknowledgments

We thank L. Bianco for histology; R. Sachidanandam for bioinformatics support; C. Rosenthal, M.-M. Yang, and S. Ray for technical assistance; and R. Dickins and other members of the Lowe laboratory for advice and critical reading of the manuscript. This work was supported by a postdoctoral fellowship from the Helen Hay Whitney Foundation (to M.T.H.), a postdoctoral training grant (to J.T.Z.), National Cancer Institute Project Grants CA13106 (to G.J.H. and S.W.L.) and CA87497 (to S.W.L.), and a generous gift from the Laurie Strauss Leukemia Foundation. D.J.B. is the recipient of a Watson School of Biological Sciences Engelhorn Fellowship, and S.W.L. is an American Association for Cancer Research–National Foundation for Cancer Research Research Professor.

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Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 101 | No. 25
June 22, 2004
PubMed: 15192153

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Submission history

Received: April 4, 2004
Published online: June 10, 2004
Published in issue: June 22, 2004

Acknowledgments

We thank L. Bianco for histology; R. Sachidanandam for bioinformatics support; C. Rosenthal, M.-M. Yang, and S. Ray for technical assistance; and R. Dickins and other members of the Lowe laboratory for advice and critical reading of the manuscript. This work was supported by a postdoctoral fellowship from the Helen Hay Whitney Foundation (to M.T.H.), a postdoctoral training grant (to J.T.Z.), National Cancer Institute Project Grants CA13106 (to G.J.H. and S.W.L.) and CA87497 (to S.W.L.), and a generous gift from the Laurie Strauss Leukemia Foundation. D.J.B. is the recipient of a Watson School of Biological Sciences Engelhorn Fellowship, and S.W.L. is an American Association for Cancer Research–National Foundation for Cancer Research Research Professor.

Authors

Affiliations

Michael T. Hemann
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724; and Genetics Program, Stony Brook University, Stony Brook, NY 11794
Jack T. Zilfou
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724; and Genetics Program, Stony Brook University, Stony Brook, NY 11794
Zhen Zhao
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724; and Genetics Program, Stony Brook University, Stony Brook, NY 11794
Darren J. Burgess
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724; and Genetics Program, Stony Brook University, Stony Brook, NY 11794
Gregory J. Hannon
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724; and Genetics Program, Stony Brook University, Stony Brook, NY 11794
Scott W. Lowe§
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724; and Genetics Program, Stony Brook University, Stony Brook, NY 11794

Notes

§
To whom correspondence should be addressed. E-mail: [email protected].
M.T.H. and J.T.Z. contributed equally to this work.
Communicated by Bruce W. Stillman, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, May 10, 2004

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    Suppression of tumorigenesis by the p53 target PUMA
    Proceedings of the National Academy of Sciences
    • Vol. 101
    • No. 25
    • pp. 9173-9513

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