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Blood, 15 October 2006, Vol. 108, No. 8, pp. 2836-2845.
Prepublished online as a Blood First Edition Paper on June 15, 2006; DOI 10.1182/blood-2006-04-016394.
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RED CELLS
MBD2 is a critical component of a methyl cytosine-binding protein complex isolated from primary erythroid cells
Evan P. Kransdorf,
Shou Zhen Wang,
Sheng Zu Zhu,
Timothy B. Langston,
Jeremy W. Rupon, and
Gordon D. Ginder
From the Massey Cancer Center, the Department of Internal Medicine, the Department of Microbiology and Immunology, the Department of Human Genetics, and the Department of Pharmacology and Toxicology, Virginia Commonwealth University School of Medicine, Richmond.
The chicken embryonic  -type globin gene,  , is a member of a small group of vertebrate genes whose developmentally regulated expression is mediated by DNA methylation. Previously, we have shown that a methyl cytosine-binding complex binds to the methylated -globin gene in vitro. We have now chromatographically purified and characterized this complex from adult chicken primary erythroid cells. Four components of the MeCP1 transcriptional repression complex were identified: MBD2, RBAP48, HDAC2, and MTA1. These 4 proteins, as well as the zinc-finger protein p66 and the chromatin remodeling factor Mi2, were found to coelute by gel-filtration analysis and pull-down assays. We conclude that these 6 proteins are components of the MeCPC. In adult erythrocytes, significant enrichment for MBD2 is seen at the inactive -globin gene by chromatin immunoprecipitation assay, whereas no enrichment is observed at the active A-globin gene, demonstrating MBD2 binds to the methylated and transcriptionally silent -globin gene in vivo. Knock-down of MBD2 resulted in up-regulation of a methylated -gene construct in mouse erythroleukemic (MEL)- cells. These results represent the first purification of a MeCP1-like complex from a primary cell source and provide support for a role for MBD2 in developmental gene regulation.
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