Abstract
Forkhead transcription factors are key participants in development and immune regulation. Here we demonstrate that absence of the gene encoding the forkhead transcription factor Foxp1 resulted in a profound defect in early B cell development. Foxp1 deficiency was associated with decreased expression of all B lineage genes in B220+ fetal liver cells as well as with a block in the transition from pro–B cell to pre–B cell involving diminished expression of recombination-activating genes 1 and 2. Foxp1 bound to the Erag enhancer and was involved in controlling variable-(diversity)-joining recombination of the gene encoding immunoglobulin heavy chain in a B cell lineage–specific way. Our results identify Foxp1 as an essential participant in the transcriptional regulatory network of B lymphopoiesis.
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Acknowledgements
We thank K. Rajewsky, S. Casola, K. Otipoby, C. Xiao, G. Galler, K. Mark Ansel and H. Liu for reagents and discussions; S. Monticelli for critical reading of the manuscript; B. Tanasa for help with the bioinformatic analyses; L. Smith for preparing genomic DNA for genotyping; C. Das and C. Schmidt for technical help; and N. Barteneva and K. Ketman for flow cytometry. Supported by the National Institutes of Health (T32 to H.H.; CA42471, AI48213 and AI44432 to A.R.; and HL071160, CA92318 and CA31534 to P.W.T.) and the Mary Betzner Morrow Centennial endowment in Molecular Genetics (P.W.T.).
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H.H. and A.R. initiated the collaboration with P.W.T. to analyze the immune phenotype of Foxp1−/− mice; B.W. and S.M. generated the Foxp1−/− mice; H.H. was responsible for all analyses of B cell development in fetal liver chimeras and in vitro pro–B cell cultures, including analysis of Foxp1 expression and chromatin immunoprecipitation assays; L.A. did EMSA and footprinting experiments under the supervision of P.W.T.; M.B. helped with mouse breeding and fetal liver collection; J.N. helped with bioinformatic analysis; and A.R. and P.W.T. provided overall supervision.
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Supplementary information
Supplementary Fig. 1
Gene targeting strategy, expression pattern of Foxp1 at different B cell developmental stages and the kinetics of peripheral B cell recovery in reconstituted RAG2-deficient recipient mice. (PDF 688 kb)
Supplementary Fig. 2
No difference in total peripheral B cell numbers between Foxp1+/+ and Foxp1+/− mice. (PDF 378 kb)
Supplementary Fig. 3
Foxp1−/− thymocyte development. (PDF 415 kb)
Supplementary Fig. 4
Expression of IL-7R on Foxp1+/− or Foxp1−/− pro-B cells and Foxp1 ChIP in thymocytes. (PDF 354 kb)
Supplementary Fig. 5
Identification of Foxp1 binding sites within KpnI-PvuII region by DNase I footprinting and EMSA. (PDF 1430 kb)
Supplementary Fig. 6
Donor cell recovery in reconstituted RAG2-deficient recipients. (PDF 410 kb)
Supplementary Table 1
Sequences of primers for Foxp1 expression and B lineage gene expression. (PDF 56 kb)
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Hu, H., Wang, B., Borde, M. et al. Foxp1 is an essential transcriptional regulator of B cell development. Nat Immunol 7, 819–826 (2006). https://doi.org/10.1038/ni1358
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DOI: https://doi.org/10.1038/ni1358
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