GATA-1-dependent transcriptional repression of GATA-2 via disruption of positive autoregulation and domain-wide chromatin remodeling

doi:10.1073/pnas.1432147100

GATA-1-dependent transcriptional repression of GATA-2 via disruption of positive autoregulation and domain-wide chromatin remodeling

^*Molecular and Cellular Pharmacology Program, Department of Pharmacology, University of Wisconsin Medical School, 383 Medical Sciences Center, 1300 University Avenue, Madison, WI 53706; and ^‡Childrens Hospital of Philadelphia, Abramson Research Center, Philadelphia, PA 19104

Edited by Stuart H. Orkin, Harvard Medical School, Boston, MA, and approved May 21, 2003 (received for review April 12, 2003)

Abstract

Interplay among GATA transcription factors is an important determinant of cell fate during hematopoiesis. Although GATA-2 regulates hematopoietic stem cell function, mechanisms controlling GATA-2 expression are undefined. Of particular interest is the repression of GATA-2, because sustained GATA-2 expression in hematopoietic stem and progenitor cells alters hematopoiesis. GATA-2 transcription is derepressed in erythroid precursors lacking GATA-1, but the underlying mechanisms are unknown. Using chromatin immunoprecipitation analysis, we show that GATA-1 binds a highly restricted upstream region of the ≈70-kb GATA-2 domain, despite >80 GATA sites throughout the domain. GATA-2 also binds this region in the absence of GATA-1. Genetic complementation studies in GATA-1-null cells showed that GATA-1 rapidly displaces GATA-2, which is coupled to transcriptional repression. GATA-1 also displaces CREB-binding protein (CBP), despite the fact that GATA-1 binds CBP in other contexts. Repression correlates with reduced histone acetylation domain-wide, but not altered methylation of histone H3 at lysine 4. The GATA factor-binding region exhibited cell-type-specific enhancer activity in transient transfection assays. We propose that GATA-1 instigates GATA-2 repression by means of disruption of positive autoregulation, followed by establishment of a domain-wide repressive chromatin structure. Such a mechanism is predicted to be critical for the control of hematopoiesis.

Footnotes

↵ § To whom correspondence should be addressed. E-mail: ehbresni{at}facstaff.wisc.edu.
↵ † J.A.G. and M.E.B. contributed equally to this work.
This paper was submitted directly (Track II) to the PNAS office.
Abbreviations: BMP-4, bone morphogenetic protein 4; CBP, CREB-binding protein; pol II, RNA polymerase II; ChIP, chromatin immunoprecipitation; ER-GATA-1, estrogen receptor hormone-binding domain fused to GATA-1; FOG-1, Friend of GATA-1; HDAC, histone deacetylase; H3-meK4, histone H3 methylated at lysine 4; HS, hypersensitive site; MEL, mouse erythroleukemia.