PMID: 8946914Nov 1, 1996Paper

A dominant interfering Myb mutant causes apoptosis in T cells

Genes & Development
D TaylorK Weston

Abstract

The c-Myb transcription factor is required for the production of most hemopoietic lineages, but information is sparse about its mode of action and the key genes it regulates. We have made an inducible dominant interfering Myb protein, by creating a chimera comprising the DNA binding domain of c-Myb, the Drosophila Engrailed repressor domain, and a modified estrogen receptor hormone binding domain. When expressed in the murine thymoma cell line EL4, activation of this mutant results in a significant proportion of the cell population undergoing apoptosis, as assessed by nuclear breakdown and DNA fragmentation, but has no apparent effect on cell-cycle progression. The apoptotic phenotype is mirrored during thymopoiesis in transgenic mice expressing dominant interfering Myb mutants; their T cells are fragile both in vivo and in vitro. Induction of the Myb dominant interfering mutant in EL4 cells correlates with down-regulation of bcl-2, but does not affect transcription of other bcl-2 family members; conversely, overexpression of bcl-2 in the transgenic mouse model rescues thymocytes from death. Analysis of the bcl-2 promoter by run-on transcription, bandshifting, and transient expression assays shows that it is a direct target of ...Continue Reading

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