Critical functions for c-Myb at three checkpoints during thymocyte development

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Abstract

The transcription factor c-Myb is expressed throughout T cell development in the thymus. However, little is understood about c-Myb function because of the embryonic lethality of traditional Myb-null mutations. Using tissue-specific deletion to abrogate c-Myb expression at distinct stages of T cell development, we identify three points at which c-Myb activity is required for normal T cell differentiation: transition through the double-negative 3 stage, survival of preselection CD4+CD8+ thymocytes, and differentiation of CD4 thymocytes. Thus, c-Myb is essential at several stages during T cell development in the thymus.

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Figure 1: Expression of Myb mRNA in the thymus and conditional targeting of the mouse Myb locus.
Figure 2: Tissue- and differentiation-specific deletion of the mouse Myb locus.
Figure 3: Impaired thymocyte development in Mybf/d LckCre and Mybf/f cwLckCre mice.
Figure 4: Differentiation and proliferation but decreased intracellular TCRβ expression in c-Myb-deficient DN3 thymocytes.
Figure 5: Inefficient V(D)J recombination at the Tcrb locus in c-Myb-deficient DN3 thymocytes.
Figure 6: c-Myb is required for DP thymocyte survival and is more important for differentiation of CD4 SP than CD8 SP cells.
Figure 7: Decreased Vα→Jα rearrangement at the Tcra locus in c-Myb-deficient DP thymocytes.

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Acknowledgements

The authors thank A. Roth (University of Cologne, Cologne, Germany) for doing the blastocyst injections; C. Goettlinger (University of Cologne) and J. Lannigan (University of Virginia, Charlottesville, Virginia) for help with cell sorting; and M. McDuffie, V. Engelhard, U. Lorenz and K. Ravichandran for critical review of the manuscript. Supported by National Institutes of Health (CA85842 to T.P.B.), Fogarty International Center (TW02297 to T.P.B.), Deutsche Forschungsgemeinschaft (SFB 243 to K.R.) and the European Union (BIO4-CT96-0077 to K.R.).

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Correspondence to Timothy P Bender.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Expression of c-Myb protein in Mybf/f lckCre thymocytes. (PDF 169 kb)

Supplementary Fig. 2

Semi-quantitative RT-PCR analysis of Rag1 and Rag2 expression in c-Myb-deficient thymocytes. (PDF 232 kb)

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Bender, T., Kremer, C., Kraus, M. et al. Critical functions for c-Myb at three checkpoints during thymocyte development. Nat Immunol 5, 721–729 (2004) doi:10.1038/ni1085

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