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p16INK4A and p19ARF act in overlapping pathways in cellular immortalization

Nature Cell Biology volume 2, pages 148–155 (2000)Cite this article

Abstract

The INK4A locus encodes two independent but overlapping genes, p16INK4A and p19ARF, and is frequently inactivated in human cancers. The unusual structure of this locus has lead to ambiguity regarding the biological role of each gene. Here we express, in primary mouse embryonic fibroblasts (MEFs), antisense RNA constructs directed specifically towards either p16INK4A or p19ARF. Such constructs induce extended lifespan in primary MEFs; this lifespan extension is reversed upon subsequent elimination of the p16INK4A or p19ARF antisense constructs. In immortal derivatives of cell lines expressing antisense p16INK4A or p19ARF RNA, growth arrest induced by recovery of p16INK4A expression is bypassed by compromising the function of the retinoblastoma protein (Rb), whereas growth arrest induced by re-expression of p19ARF is overcome only by simultaneous inactivation of both the Rb and the p53 pathways. Thus, the physically overlapping p16INK4A and p19ARF genes act in partly overlapping pathways.

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Figure 1: Inactivation of p19ARF and p16INK4A.
Figure 2: Inhibition of p16INK4A and p19ARF is reversible.
Figure 3: Overlapping functions of p16INK4A and p19 ARF.
Figure 4: MDM2 contributes to immortalization of MEFs.
Figure 5: E2F-1 can overcome the growth arrest induced by p19ARF in p53-null cells.
Figure 6: Analysis of p16INK4A and p19ARF messenger RNA expression in p53–/– immortalized clones.
Figure 7: Inactivation of p16INK4A or p19ARF cooperates with loss of p53 in immortalization.
Figure 8: Model of gene function in immortalization of mouse cells.

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Acknowledgements

We thank H. Brady for providing p53–/– MEFs; A. Badley for MDM2–/– p53–/– MEFs; and M. Serrano and G. Hannon for critical reading of the manuscript. A.C. is a recipient of an EMBO long-term fellowship. J.D.H. is supported by a fellowship from the Leukaemia Research Fund. J.D.H. and A.C. were also supported by the Cancer Research Campaign. D.H.B. is supported by the Hugh and Catherine Stevenson Fund and Cancer Research Campaign.

Correspondence and requests for materials should be addressed to D.H.B.

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  1. Amancio Carnero & David H. Beach

    Present address: Institute of Child Health, UCL, Cruciform Building, Gower Street, London, WC1E 6BT, UK

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  1. Institute of Child Health, 30 Guilford Street, London, WC1 1EH, UK

    Amancio Carnero, James D. Hudson, Cathy M. Price & David H. Beach

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Carnero, A., Hudson, J., Price, C. et al. p16INK4A and p19ARF act in overlapping pathways in cellular immortalization. Nat Cell Biol 2, 148–155 (2000). https://doi.org/10.1038/35004020

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