Coreceptor expression is tightly regulated during thymocyte development. Deletion of specific Cd8 enhancers leads to variegated expression of CD8αβ heterodimers in double-positive thymocytes. Here we show CD8 variegation is correlated with an epigenetic 'off' state, linking Cd8 enhancer function with chromatin remodeling of the adjacent genes Cd8a and Cd8b1 (Cd8). The zinc finger protein MAZR bound the Cd8 enhancer and interacted with the nuclear receptor corepressor N-CoR complex in double-negative thymocytes. MAZR was downregulated in double-positive and CD8 single-positive thymocytes. 'Enforced' expression of MAZR led to impaired Cd8 activation and variegated CD8 expression. Our results demonstrate epigenetic control of the Cd8 loci and identify MAZR as an important regulator of Cd8 expression.
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We thank V.J. Bardwell for N-CoR expression constructs; E. Pfeiffer and E. Selzev for help with irradiation; and S. Sakaguchi, P. Kinross and R. Herbst for critical reading of the manuscript. Supported by the Austrian Science Fund (P14261 and P16708), the START Program (Project Y-163) of the Austrian Ministry of Education, Science and Culture, the K-plus Competence Center Biomolecular Therapeutics, the Federal Bank of Austria (Project 10530), the Austrian Program for Advanced Research and Technology of the Austrian Academy of Sciences (W.E.) and the National Institutes of Health (HD39454 and HD18184 to C.B.W.).
The authors declare no competing financial interests.
Loss of Dnmt1 leads to a partial reactivation of CD8 expression in variegated DP cells. (PDF 80 kb)
Map of the clone isolated in the YOH screen and localization of the bait RE-1 sequence. (PDF 123 kb)
Specificity of the anti-MAZR serum. (PDF 84 kb)
Retroviral constructs used for the transduction of hematopoietic stem cells. (PDF 95 kb)
CD8-negative DP thymocytes develop upon forced expression of MAZR in Tcra/ thymocytes. (PDF 140 kb)
PCR primers used for bisulfite sequencing analysis. (PDF 12 kb)
PCR primers used for ChIP assays. (PDF 16 kb)
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Bilic, I., Koesters, C., Unger, B. et al. Negative regulation of CD8 expression via Cd8 enhancer–mediated recruitment of the zinc finger protein MAZR. Nat Immunol 7, 392–400 (2006). https://doi.org/10.1038/ni1311
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