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A conserved motif within RAP1 has diversified roles in telomere protection and regulation in different organisms

Nature Structural & Molecular Biology volume 18, pages 213–221 (2011)Cite this article

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Abstract

Repressor activator protein 1 (RAP1) is the most highly conserved telomere protein. It is involved in protecting chromosome ends in fission yeast and promoting gene silencing in Saccharomyces cerevisiae, whereas it represses homology-directed recombination at telomeres in mammals. To understand how RAP1 has such diverse functions at telomeres, we solved the crystal or solution structures of the RAP1 C-terminal (RCT) domains of RAP1 from multiple organisms in complex with their respective protein-binding partners. Our analysis establishes RAP1RCT as an evolutionarily conserved protein-protein interaction module. In mammalian and fission yeast cells, this module interacts with TRF2 and Taz1, respectively, targeting RAP1 to chromosome ends for telomere protection. In contrast, S. cerevisiae RAP1 uses its RCT domain to recruit Sir3 to telomeres to mediate gene silencing. Together, our results show that, depending on the organism, the evolutionarily conserved RAP1 RCT motif has diverse functional roles at telomeres.

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Figure 1: Structure of the human TRF2RBM–RAP1RCT complex.
Figure 2: Crystallographic and mutational analyses of TRF2RBM–RAP1RCT interaction.
Figure 3: RAP1 is required for telomere end protection.
Figure 4: Structure of fission yeast Taz1RBM–SpRap1RCT complex.
Figure 5: Mutational analyses of Taz1-SpRap1 interaction.
Figure 6: Structure of budding yeast Sir3RBM–ScRap1RCT complex and its role in telomeric silencing.

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Acknowledgements

We thank J. Karlseder (Salk Institute for Biological Studies), L. Rudolph (University of Ulm) and M. Tarsounas (Cancer Research UK and University of Oxford) for providing valuable reagents. We thank F. Ishikawa (Kyoto University) for strains and Y. Ogiyama and K. Ishii (Osaka University) for technical support on PFGE. M.L. acknowledges financial support from the US National Institutes of Health (RO1 GM083015), the American Cancer Society and the Sidney Kimmel Foundation. M.L. is a Howard Hughes Medical Institute Early Career Scientist. S.C. acknowledges financial support from the US National Institute on Aging (RO1 AG028888), the US National Cancer Institute (RO1 CA129037), the Welch Foundation, the Susan G. Komen Race for the Cure Foundation, the Abraham and Phyllis Katz Foundation and the Michael Kadoorie Cancer Genetic Research Program. J.K. was supported by grants from Osaka University Life Science Young Independent Researcher Support Program (Japan Science and Technology Agency), the Japanese Ministry of Education, Culture, Sports, Science and Technology, Inamori Foundation, Astellas Foundation for Research on Metabolic Disorders and Takeda Science Foundation. Work in the laboratory of D.S. was supported by a grant from the Swiss National Fund (grant 31003A116716), by the Swiss National Fund Frontiers in Genetics NCCR program and by the Canton of Geneva. Use of Life Sciences Collaborative Access Team Sector 21 was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (grant 085P1000817). Use of the Advanced Photon Source (APS) was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences (contract no. DE-AC02-06CH11357).

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  1. Rekha Rai & Sandy Chang

    Present address: Present address: Department of Laboratory Medicine, Yale University School of Medicine, New Haven, Connecticut, USA.,

  2. Yong Chen and Rekha Rai: These authors contributed equally to this work.

Authors and Affiliations

  1. Howard Hughes Medical Institute, University of Michigan Medical School, Ann Arbor, Michigan, USA

    Yong Chen, Yuting Yang, Feng Wang & Ming Lei

  2. Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan, USA

    Yong Chen, Yuting Yang, Feng Wang & Ming Lei

  3. Department of Genetics, M.D. Anderson Cancer Center, Houston, Texas, USA

    Rekha Rai, Hong Zheng & Sandy Chang

  4. State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

    Zi-Ren Zhou & Hong-Yu Hu

  5. Institute for Protein Research, Osaka University, Suita, Osaka, Japan

    Junko Kanoh & Hisayo Tsujii

  6. Department of Molecular Biology and National Centre of Competence in Research (NCCR) Frontiers in Genetics Program, University of Geneva, Geneva, Switzerland

    Cyril Ribeyre, Pascal Damay & David Shore

  7. Graduate School of Frontier Biosciences, Osaka University, Osaka, Suita, Japan

    Yasushi Hiraoka

  8. Department of Hematopathology, M.D. Anderson Cancer Center, Houston, Texas, USA

    Sandy Chang

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Contributions

M.L., S.C., H.H., J.K. and D.S. designed the experiments. Y.C., R.R., Z.Z., C.R., Y.Y., H.Z., P.D., F.W. and H.T. conducted the experiments. M.L., S.C., H.H., J.K., D.S. and Y.H. analyzed the data. M.L. wrote the paper.

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Correspondence to Sandy Chang or Ming Lei.

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Chen, Y., Rai, R., Zhou, ZR. et al. A conserved motif within RAP1 has diversified roles in telomere protection and regulation in different organisms. Nat Struct Mol Biol 18, 213–221 (2011). https://doi.org/10.1038/nsmb.1974

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