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A systematic RNAi synthetic interaction screen reveals a link between p53 and snoRNP assembly

Nature Cell Biology volume 13, pages 809818 (2011) | Download Citation

Abstract

TP53(tumour protein 53) is one of the most frequently mutated genes in human cancer and its role during cellular transformation has been studied extensively. However, the homeostatic functions of p53 are less well understood. Here, we explore the molecular dependency network of TP53 through an RNAi-mediated synthetic interaction screen employing two HCT116 isogenic cell lines and a genome-scale endoribonuclease-prepared short interfering RNA library. We identify a variety of TP53 synthetic interactions unmasking the complex connections of p53 to cellular physiology and growth control. Molecular dissection of the TP53 synthetic interaction with UNRIP indicates an enhanced dependency of TP53-negative cells on small nucleolar ribonucleoprotein (snoRNP) assembly. This dependency is mediated by the snoRNP chaperone gene NOLC1 (also known as NOPP140), which we identify as a physiological p53 target gene. This unanticipated function of TP53 in snoRNP assembly highlights the potential of RNAi-mediated synthetic interaction screens to dissect molecular pathways of tumour suppressor genes.

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Acknowledgements

We would like to thank all members of the Buchholz laboratory for discussions and sharing reagents. We thank A. K. Heninger for help with carrying out the screen, V. Surendranath for esiRNA design, M. Theis, S. Rose and A. Weise for esiRNA production and I. Poser for assistance with BAC cell line generation. This work was supported by the Max Planck Society, the German Federal Ministry of Education and Research grants Go-Bio (0315105), DiGtoP (01GS0859) and the DFG grant SFB655.

Author information

Author notes

    • Nina C. Hubner

    Present address: Universitair Medisch Centrum Utrecht, 3508 AB Utrecht, The Netherlands

    • Magno Junqueira

    Present address: Brazilian Center for Protein Research, Department of Cell Biology, University of Brasilia, 70910-900 Brasilia, DF, Brazil

Affiliations

  1. University of Technology Dresden, University Hospital and Medical Faculty Carl Gustav Carus, Department of Medical Systems Biology, Fetscherstraße 74, D-01307 Dresden, Germany

    • Dragomir B. Krastev
    • , Maciej Paszkowski-Rogacz
    •  & Frank Buchholz
  2. Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany

    • Dragomir B. Krastev
    • , Mikolaj Slabicki
    • , Maciej Paszkowski-Rogacz
    • , Magno Junqueira
    • , Andrej Shevchenko
    • , Karla M. Neugebauer
    •  & Frank Buchholz
  3. Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany

    • Nina C. Hubner
    •  & Matthias Mann

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Contributions

D.B.K., M.S., N.C.H., M.J. and K.M.N. carried out experiments, M.P-R. analysed data, A.S., M.M. and F.B. planned the project and D.B.K. and F.B. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Frank Buchholz.

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https://doi.org/10.1038/ncb2264

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