Conditional Knockout Mice Reveal Distinct Functions for the Global Transcriptional Coactivators CBP and p300 in T-Cell Development

Abstract

The global transcriptional coactivators CREB-binding protein (CBP) and the closely related p300 interact with over 312 proteins, making them among the most heavily connected hubs in the known mammalian protein-protein interactome. It is largely uncertain, however, if these interactions are important in specific cell lineages of adult animals, as homozygous null mutations in either CBP or p300 result in early embryonic lethality in mice. Here we describe a Cre/LoxP conditional p300 null allele (p300 ^flox) that allows for the temporal and tissue-specific inactivation of p300. We used mice carrying p300 ^flox and a CBP conditional knockout allele (CBP ^flox) in conjunction with an Lck-Cre transgene to delete CBP and p300 starting at the CD4⁻ CD8⁻ double-negative thymocyte stage of T-cell development. Loss of either p300 or CBP led to a decrease in CD4⁺ CD8⁺ double-positive thymocytes, but an increase in the percentage of CD8⁺ single-positive thymocytes seen in CBP mutant mice was not observed in p300 mutants. T cells completely lacking both CBP and p300 did not develop normally and were nonexistent or very rare in the periphery, however. T cells lacking CBP or p300 had reduced tumor necrosis factor alpha gene expression in response to phorbol ester and ionophore, while signal-responsive gene expression in CBP- or p300-deficient macrophages was largely intact. Thus, CBP and p300 each supply a surprising degree of redundant coactivation capacity in T cells and macrophages, although each gene has also unique properties in thymocyte development.

We thank S. Lerach and C. Barlow for excellent technical assistance; the Transgenic core; R. Cross, J. Gatewood, J. Smith, and the Mouse Immunophenotyping core; R. Ashmun and the Flow Cytometry and Cell Sorting Shared Resource; the Animal Resource Center facilities at SJCRH; M. Biery, N. Craig, G. Martin, and S. Fiering for plasmids; and J. Opferman for helpful comments. The Hartwell Center at SJCRH provided oligonucleotides and DNA sequencing.

This work was supported by NCI grant RO1 CA076385 (to P.K.B.), the Cancer Center (CORE) support grant P30 CA021765, and by the American Lebanese Syrian Associated Charities of St. Jude Children's Research Hospital.