p53, the Cellular Gatekeeper for Growth and Division

As the tale of p53 unfolds, it becomes ever more intriguing. Although our understanding of the critical and complex roles played by p53 is progressing rapidly, new findings continue to pose new paradoxes. Here we present some of these recent advances in p53 research and discuss how they either shed light on or add to the complexities of p53. Therefore, we only briefly summarize some of the key developments leading to our current state of knowledge. For further information, the reader is referred to several excellent reviews that have focused on p53 research (see Donehower and Bradley 1993; Levine 1993; Greenblatt et al. 1994; Oren 1994; Prives 1994; Kinzler and Vogelstein 1996).

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p53: puzzle and paradigm.

Tumor spectrum analysis in p53-mutant mice.

In the past fifteen years, it has become apparent that tumour-associated p53 mutations can provoke activities that are different to those resulting from simply loss of wild-type tumour-suppressing p53 function. Many of these mutant p53 proteins acquire oncogenic properties that enable them to promote invasion, metastasis, proliferation and cell survival. Here we highlight some of the emerging molecular mechanisms through which mutant p53 proteins can exert these oncogenic functions.

p53 mutations in cancer

The crucial differences between normal cells and cancer cells stem from discrete changes in specific genes controlling proliferation and tissue homeostasis. Over 100 such cancer-related genes have been discovered, several of which are implicated in the natural history of human cancer because they are consistently found to be mutated in tumors. The p53 tumor-suppressor gene is the most striking example because it is mutated in about half of almost all types of cancer arising from a wide spectrum of tissues. Other tumor-suppressor genes important in human cancers, such as adenomatous polyposis coli, Wilms' tumor type 1, and neurofibromatosis type 1, . . .

Clinical implications of the p53 tumor-suppressor gene

We report that the expression of murine or human mutant p53 proteins in cells with no endogenous p53 proteins confers new or additional phenotypes upon these cells. Mutant p53 proteins expressed in cell lines lacking p53 resulted in either enhanced tumorigenic potential in nude mice ((10)3 cells) or enhanced plating efficiency in agar cell culture (human SAOS-2 cells). Also, mutant human p53 alleles, unlike the wild-type p53 protein, could also enhance the expression of a test gene regulated by the multi-drug resistance enhancer-promoter element. These data demonstrate a gain of function associated with p53 mutations in addition to the loss of function shown previously to be associated with mutations in this tumour suppressor gene.

Gain of function mutations in p53.

Under conditions in which cytoplasmic accumulation of HeLa cell mRNAs has been blocked by adenovirus infection, hsp70 family mRNAs are transported from the nucleus to the cytoplasm at near normal efficiency subsequent to heat shock. Heat shock does not reverse the general virus-induced block to host cell mRNA transport. The heat shock mRNAs are translated within the cytoplasm of the infected cell but at substantially reduced efficiency compared with that of uninfected cells. Thus, the hsp70 family of mRNAs can escape the transport block but not the translational block instituted late after adenovirus infection. The beta-tubulin gene family is induced by the viral E1A gene after infection, and its mRNAs also accumulate in the cytoplasmic compartment. Given these two examples, it seems likely that the process of transcriptional induction allows the resulting mRNA to escape the viral block of transport.

Induced heat shock mRNAs escape the nucleocytoplasmic transport block in adenovirus-infected HeLa cells.

Many diverse tumors contain cells that select for mutations at the p53 gene locus. This appears to be the case because the p53 gene product can act as a negative regulator of cell division or a tumor suppressor. These mutations then eliminate this activity of the p53 gene product. The simian virus 40 (SV40) large T antigen binds to p53 and acts as an oncogene to promote cellular transformation and initiate tumors. If the binding of T antigen to the p53 protein inactivated its tumor suppressor activity, there would be no selection pressure for p53 mutants to appear in tumors. To test this idea, transgenic mice that carried and expressed the SV40 large T-antigen gene were created. Expression of the T antigen was directed to the liver, using the albumin promoter, and the choroid plexus, using the SV40 enhancer-promoter. A large number of papillomas (indicated in parentheses) of the choroid plexus (14), hepatocellular carcinomas (5), liver adenomas (10), and tumors of clear-cell foci (5) were examined for mutant and wild-type p53 genes and gene products. In all cases, the tumor extracts contained readily detectable T-antigen-p53 protein complexes. A monoclonal antibody specifically recognizing the wild-type p53 protein (PAb246) reacted with p53 in every tumor extract. A monoclonal antibody specifically recognizing mutant forms of the p53 protein (PAb240) failed to detect p53 antigens in these extracts. Finally, p53 partial cDNAs were sequenced across the regions of common mutations in this gene, and in every case only the wild-type sequence was detected. These results strongly support the hypothesis that T antigen inactivates the wild-type p53 tumor-suppressing activity and there is no need to select for mutations at the p53 locus.

p53 mutations are not selected for in simian virus 40 T-antigen-induced tumors from transgenic mice.

All mRNAs encoded by the adenovirus major late transcription unit share a common 5' noncoding region, 200 nucleotides in length, termed the tripartite leader sequence. To assess function of the tripartite leader, recombinant viruses were prepared which carried either a bona fide herpes simplex virus thymidine kinase gene or a modified thymidine kinase gene whose normal 5' noncoding domain was replaced with the adenovirus leader sequence. The tripartite leader simultaneously decreased the nuclear half-life and increased the cytoplasmic half-life of the thymidine kinase-specific mRNA. The tripartite leader stabilized the non-adenovirus mRNA only within the environment of an adenovirus-infected cell during the late phase of the infectious cycle.

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M Moore

Papers

Gain of function mutations in p53.

Induced heat shock mRNAs escape the nucleocytoplasmic transport block in adenovirus-infected HeLa cells.

p53 mutations are not selected for in simian virus 40 T-antigen-induced tumors from transgenic mice.

The adenovirus tripartite leader sequence can alter nuclear and cytoplasmic metabolism of a non-adenovirus mRNA within infected cells

adenovirus-infected HeLa cells. nucleocytoplasmic transport block in Induced heat shock mRNAs escape the