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Regulation of hematopoietic stem cell differentiation by a single ubiquitin ligase–substrate complex

Nature Immunology volume 11, pages 207–215 (2010)Cite this article

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Abstract

Hematopoietic stem cell (HSC) differentiation is regulated by cell-intrinsic and cell-extrinsic cues. In addition to transcriptional regulation, post-translational regulation may also control HSC differentiation. To test this hypothesis, we visualized the ubiquitin-regulated protein stability of a single transcription factor, c-Myc. The stability of c-Myc protein was indicative of HSC quiescence, and c-Myc protein abundance was controlled by the ubiquitin ligase Fbw7. Fine changes in the stability of c-Myc protein regulated the HSC gene-expression signature. Using whole-genome genomic approaches, we identified specific regulators of HSC function directly controlled by c-Myc binding; however, adult HSCs and embryonic stem cells sensed and interpreted c-Myc-regulated gene expression in distinct ways. Our studies show that a ubiquitin ligase–substrate pair can orchestrate the molecular program of HSC differentiation.

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Figure 1: Abundance of c-Myc protein in early hematopoiesis.
Figure 2: Correlation between the amount of c-Myc protein and loss of HSC self-renewal.
Figure 3: Fbw7 controls the stability of c-Myc protein in HSCs.
Figure 4: The abundance of c-Myc protein directly controls the molecular program of stem cell differentiation, cell-cycle entry and self-renewal.
Figure 5: Genes overexpressed in c-Myc-EGFPhi cells are directly bound by the c-Myc transcription factor.
Figure 6: The role of the c-Myc–Fbw7 interaction in fetal liver stem cells and progenitor cells.
Figure 7: Expression patterns of Fbw7 and c-Myc in mouse ESCs.
Figure 8: Fbw7 is dispensable for the self-renewal of mouse ESCs.

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Acknowledgements

We thank the members of the Aifantis Lab and specifically B. King for advice as well as time and effort spent on this manuscript; P. Lopez and the NYU Flow Cytometry Facility for cell sorting; the NYU Cancer Institute Genomics Facility for help with microarray processing; M. Gostissa (Harvard University) and F. Alt (Harvard University) for Mycflox mice; and I. Lemischka and his laboratory (Mount Sinai) for the Nanog-GFP line and technical advice. Supported by the National Institutes of Health (RO1CA133379, RO1CA105129, R21CA141399, R56AI070310 and P30CA016087 to I.A.; RO1AI41428 and RO1AI072039 to B.P.S.; and R01CA120196 to A.F.), the American Cancer Society (RSG0806801 to I.A.), the Edward Mallinckrodt Jr. Foundation, the Irma T. Hirschl Trust, the Alex's Lemonade Stand Foundation (I.A.), the Alexander von Humboldt Foundation (B.A.-O.), the NYU Hematology/Oncology Program (S.M.B.), an NYU Molecular Oncology and Immunology Training Grant (5T32CA009161 to K.C.), the Leukemia & Lymphoma Society (I.A. and A.F.) and the Howard Hughes Medical Institute (I.A.).

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Authors and Affiliations

  1. Howard Hughes Medical Institute and Department of Pathology, New York, New York, USA

    Linsey Reavie, Kelly Crusio, Beatriz Aranda-Orgilles, Shannon M Buckley, Benjamin Thompson, Eugine Lee, Jie Gao & Iannis Aifantis

  2. New York University (NYU) Cancer Institute and Helen & Martin S. Kimmel Stem Cell Center, NYU School of Medicine, New York, New York, USA

    Linsey Reavie, Kelly Crusio, Beatriz Aranda-Orgilles, Shannon M Buckley, Benjamin Thompson, Eugine Lee, Jie Gao, Jiri Zavadil & Iannis Aifantis

  3. Institute for Cancer Genetics, Columbia University, New York, New York, USA

    Giusy Della Gatta, Teresa Palomero & Adolfo Ferrando

  4. Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri, USA

    Andrea L Bredemeyer, Beth A Helmink & Barry P Sleckman

  5. Center for Health Informatics and Bioinformatics, NYU School of Medicine, New York, New York, USA

    Jiri Zavadil

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Contributions

L.R. did most of the experiments and participated in preparing the manuscript; G.D.G., T.P., A.F. and B.A.-O. designed and did the ChIP-plus-microarray and ChIP experiments; K.C., E.L. and B.T. did the ESC experiments; B.P.S., B.T., A.L.B. and B.A.H. generated and did initial studies with the c-Myc–eGFP mice; J.Z. analyzed microarray data; and I.A. designed the study and prepared the manuscript.

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Correspondence to Iannis Aifantis.

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Reavie, L., Gatta, G., Crusio, K. et al. Regulation of hematopoietic stem cell differentiation by a single ubiquitin ligase–substrate complex. Nat Immunol 11, 207–215 (2010). https://doi.org/10.1038/ni.1839

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