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A unique Oct4 interface is crucial for reprogramming to pluripotency

Nature Cell Biology volume 15, pages 295–301 (2013)Cite this article

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Abstract

Terminally differentiated cells can be reprogrammed to pluripotency by the forced expression of Oct4, Sox2, Klf4 and c-Myc1,2. However, it remains unknown how this leads to the multitude of epigenetic changes observed during the reprogramming process. Interestingly, Oct4 is the only factor that cannot be replaced by other members of the same family to induce pluripotency3,4,5. To understand the unique role of Oct4 in reprogramming, we determined the structure of its POU domain bound to DNA. We show that the linker between the two DNA-binding domains is structured as an α-helix and exposed to the protein’s surface, in contrast to the unstructured linker of Oct1. Point mutations in this α-helix alter or abolish the reprogramming activity of Oct4, but do not affect its other fundamental properties. On the basis of mass spectrometry studies of the interactome of wild-type and mutant Oct4, we propose that the linker functions as a protein–protein interaction interface and plays a crucial role during reprogramming by recruiting key epigenetic players to Oct4 target genes. Thus, we provide molecular insights to explain how Oct4 contributes to the reprogramming process.

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Figure 1: The crystallographic structure of the Oct4POU:PORE complex.
Figure 2: The effect of mutations in the Oct4 linker on reprogramming activity.
Figure 3: Characterization of alanine mutations on the linker segment.
Figure 4: Surface view of the Oct4POU:PORE complex.

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Acknowledgements

X-ray diffraction experiments were performed on the ID23-1 beamline at the European Synchrotron Radiation Facility (ESRF), Grenoble, and X12 and X13 at EMBL Hamburg, DESY. We are grateful to E. Fioravanti of the ESRF for providing assistance in using beamline ID23-1. We thank G. Bourenkov for invaluable assistance in crystallographic problem resolution and data collection. We thank A. Nolte for help with mass spectrometry measurements. We also thank R. Grindberg for critically reading, and A. Malapetsas and J. Bruder for editing the manuscript. We acknowledge the financial support from DFG (SPP1356 Pluripotency and Cellular Reprogramming priority program) AJ DFG SI 1695/1-2 and AJ DFG CO 975/1.

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  1. Daniel Esch and Juha Vahokoski: These authors contributed equally to this work

Authors and Affiliations

  1. Department for Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, Münster D-48149, Germany

    Daniel Esch, Vlad Cojocaru, Hermann vom Bruch, Dong Han, Hannes C. A. Drexler, Marcos J. Araúzo-Bravo & Hans R. Schöler

  2. EMBL-Hamburg, Notkestrasse 85, Hamburg D-22603, Germany

    Juha Vahokoski, Matthew R. Groves, Vivian Pogenberg & Matthias Wilmanns

  3. Laboratory for Structural Biochemistry, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore

    Calista K. L. Ng & Ralf Jauch

  4. School of Biological Sciences, Nanyang Technological University, Singapore 639798, Singapore

    Calista K. L. Ng

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Contributions

D.E. and J.V. performed the experiments and wrote the manuscript; J.V., M.R.G. and V.P. collected and analysed the crystallographic data; V.C. performed the comparative modelling experiments and wrote the manuscript; H.v.B. performed the cycloheximide experiments; H.C.A.D. performed the MS experiments; M.J.A-B. analysed the proteomic data sets; D.H. performed the rescue experiments; and C.N. assessed the heterodimer formation ability of Oct4 and Sox2. All authors commented on the manuscript; and R.J., M.W. and H.R.S. as co-senior authors supervised the project and assisted in writing the manuscript.

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Esch, D., Vahokoski, J., Groves, M. et al. A unique Oct4 interface is crucial for reprogramming to pluripotency. Nat Cell Biol 15, 295–301 (2013). https://doi.org/10.1038/ncb2680

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