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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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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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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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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DOI: https://doi.org/10.1038/ni.1839
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