CD8 Coreceptor Extinction in Signaled CD4⁺CD8⁺ Thymocytes: Coordinate Roles for Both Transcriptional and Posttranscriptional Regulatory Mechanisms in Developing Thymocytes

Abstract

T-cell development in the thymus is characterized by changing expression patterns of CD4 and CD8 coreceptor molecules and by changes in CD4 and CD8 gene transcription. In response to T-cell receptor (TCR) signals, thymocytes progress through developmental transitions, such as conversion of CD4⁺CD8⁺ (double-positive [DP]) thymocytes into intermediate CD4⁺CD8⁻thymocytes, that appear to require more-rapid changes in coreceptor expression than can be accomplished by transcriptional regulation alone. Consequently, we considered the possibility that TCR stimulation of DP thymocytes not only affects coreceptor gene transcription but also affects coreceptor RNA stability. Indeed, we found that TCR signals in DP thymocytes rapidly destabilized preexisting CD4 and CD8 coreceptor RNAs, resulting in their rapid elimination. Destabilization of coreceptor RNA was shown for CD8α to be dependent on target sequences in the noncoding region of the RNA. TCR signals also differentially affected coreceptor gene transcription in DP thymocytes, terminating CD8α gene transcription but only transiently reducing CD4 gene transcription. Thus, posttranscriptional and transcriptional regulatory mechanisms act coordinately in signaled DP thymocytes to promote the rapid conversion of these cells into intermediate CD4⁺CD8⁻ thymocytes. We suggest that destabilization of preexisting coreceptor RNAs is a mechanism by which coreceptor expression in developing thymocytes is rapidly altered at critical points in the differentiation of these cells.

ACKNOWLEDGMENTS

We are grateful to B. J. Fowlkes and Ellen Robey for providing CD8α.1-transgenic mice; Remy Bosselut, Richard Hodes, and Dinah Singer for critically reading the manuscript; Larry Granger and Tony Adams for expert flow analysis and electronic cell sorting; and Yousuke Takahama for advice on nuclear run-on assays.