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Interaction between ubiquitin–protein ligase SCFSKP2 and E2F-1 underlies the regulation of E2F-1 degradation

Nature Cell Biology volume 1, pages 14–19 (1999)Cite this article

Abstract

The transcription factor E2F-1 is important in the control of cell proliferation. Its activity must be tightly regulated in a cell-cycle-dependent manner to enable programs of gene expression to be coupled closely with cell-cycle position. Here we show that, following its accumulation in the late G1 phase of the cell cycle, E2F-1 is rapidly degraded in S/G2 phase. This event is linked to a specific interaction of E2F-1 with the F-box-containing protein p45SKP2, which is the cell-cycle-regulated component of the ubiquitin–protein ligase SCFSKP2 that recognizes substrates for this ligase. Disruption of the interaction between E2F-1 and p45SKP2 results in a reduction in ubiquitination of E2F-1 and the stabilization and accumulation of transcriptionally active E2F-1 protein. These results indicate that an SCFSKP2-dependent ubiquitination pathway may be involved in the downregulation of E2F-1 activity in the S/G2 phase of the cell cycle, and suggest a link between SCFSKP2 and cell-cycle-dependent gene control.

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Figure 1: p45 SKP2 associates with a ubiquitination-promoting activity in vitro.
Figure 2: Cell-cycle regulation of E2F-1 protein accumulation and proteasome-dependent degradation of E2F-1 in S/G2 phase.
Figure 3: Formation of p45 SKP2 , CUL-1 and E2F-1 complexes in vivo.
Figure 4: Identification of a segment of E2F-1 that is sufficient for its binding to p45 SKP2 and CUL-1 in vitro.
Figure 5: The p45 SKP2 -binding domain of E2F-1 is required for efficient ubiquitination of E2F-1 and its presence correlates with instability in vivo.
Figure 6: Effects of uncoupling E2F-1 from SCF SKP2 -dependent ubiquitination on the DNA-binding function and S-phase-promoting activity of E2F-1 in retrovirally infected NIH3T3 cells.

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Acknowledgements

We thank the members of our laboratory for many helpful discussions; H. Angliker for sequencing and P. Müller for synthesis of oligonucleotides; D. Beach for p45SKP2 and p19SKP1 cDNAs and for the GST–p45SKP2 baculovirus; A. Pause and R. Klausner for HA-tagged CUL-1 plasmid; D. Bohmann for the HA–ubiquitin plasmid; E. Harlow for pCMV–CDC2; and B. Amati, R. Eckner, P. Caroni, N. Hynes, P. Matthias and members of our laboratory for careful reading of the manuscript. W. K. is a START fellow and is supported by the Swiss National Science Foundation.

Correspondence and requests for materials should be addressed to W. K.

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Authors and Affiliations

  1. Friedrich Miescher Institut, Maulbeerstrasse 66, Basel, CH-4058, Switzerland

    A. Marti & C. Wirbelauer

  2. Deutsches Krebsforschungszentrum, Heidelberg, D-69017, Germany

    M. Scheffner

  3. W. Krek

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Marti, A., Wirbelauer, C., Scheffner, M. et al. Interaction between ubiquitin–protein ligase SCFSKP2 and E2F-1 underlies the regulation of E2F-1 degradation. Nat Cell Biol 1, 14–19 (1999). https://doi.org/10.1038/8984

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