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Published August 8, 2006 | Supplemental Material + Published + Erratum
Journal Article Open

Notch/Delta signaling constrains reengineering of pro-T cells by PU.1

Abstract

PU.1 is essential for early stages of mouse T cell development but antagonizes it if expressed constitutively. Two separable mechanisms are involved: attenuation and diversion. Dysregulated PU.1 expression inhibits pro-T cell survival, proliferation, and passage through β-selection by blocking essential T cell transcription factors, signaling molecules, and Rag gene expression, which expression of a rearranged T cell antigen receptor transgene cannot rescue. However, Bcl2 transgenic cells are protected from this attenuation and may even undergo β-selection, as shown by PU.1 transduction of defined subsets of Bcl2 transgenic fetal thymocytes with differentiation in OP9-DL1 and OP9 control cultures. The outcome of PU.1 expression in these cells depends on Notch/Delta signaling. PU.1 can efficiently divert thymocytes toward a myeloid-like state with multigene regulatory changes, but Notch/Delta signaling vetoes diversion. Gene expression analysis distinguishes sets of critical T lineage regulatory genes with different combinatorial responses to PU.1 and Notch/Delta signals, suggesting particular importance for inhibition of E proteins, Myb, and/or Gfi1 (growth factor independence 1) in diversion. However, Notch signaling only protects against diversion of cells that have undergone T lineage specification after Thy-1 and CD25 up-regulation. The results imply that in T cell precursors, Notch/Delta signaling normally acts to modulate and channel PU.1 transcriptional activities during the stages from T lineage specification until commitment.

Additional Information

© 2006 The National Academy of Sciences of the USA. Edited by Max D. Cooper, University of Alabama, Birmingham, AL, and approved June 19, 2006 (received for review February 11, 2006). Published online on July 31, 2006, 10.1073/pnas.0601188103. We thank Thomas Graf for discussion of unpublished results; J.-C. Zúñiga-Pflücker (University of Toronto, Toronto, Canada) for OP9 cell lines; Eric Davidson for insightful criticism of the manuscript and kind permission to use the ABI7900HT sequence detector; E.-S. David and C. C. Tydell (both at the California Institute of Technology) for PCR primers and help with quantitative PCR; Robert Butler for fine technical support; and Robin Condie, Ruben Bayon, Lorena Del Carmen Sandoval, Natasha Bouey, and Sherwin Chen for excellent care of mice. This work was supported by U.S. Public Health Service Grants R01 CA90233 and R01 CA98925 and by the DNA Sequencer Royalty Fund at California Institute of Technology. C.B.F. and D.D.S.-A. contributed equally to this work. Author contributions: D.D.S.-A., T.T., M.A.Y., and E.V.R. designed research; C.B.F., D.D.S.-A., I.P., T.T., A.H.W., M.A.Y., and S.L.A. performed research; C.B.F., D.D.S.-A., I.P., T.T., S.L.A., R.A.D., and E.V.R. analyzed data; E.V.R. wrote the paper; and A.H.W. provided crucial background studies. Conflict of interest statement: No conflicts declared. This paper was submitted directly (Track II) to the PNAS office. Correction. PNAS, September 19, 2006, 103(38):14255.

Errata

Correction for Franco et al., Notch/Delta signaling constrains reengineering of pro-T cells by PU.1 Proceedings of the National Academy of Sciences Sep 2006, 103 (38) 14255; DOI: 10.1073/pnas.0607045103

Attached Files

Published - FRApnas06.pdf

Supplemental Material - FRApnas06fig10.pdf

Supplemental Material - FRApnas06fig11.pdf

Supplemental Material - FRApnas06fig5.pdf

Supplemental Material - FRApnas06fig6.pdf

Supplemental Material - FRApnas06fig7.pdf

Supplemental Material - FRApnas06fig8.pdf

Supplemental Material - FRApnas06fig9.pdf

Supplemental Material - FRApnas06methods.pdf

Supplemental Material - FRApnas06table.pdf

Erratum - FRApnas06corr.pdf

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Additional details

Created:
August 22, 2023
Modified:
October 23, 2023