Context-dependent function of "GATA switch" sites in vivo.

Master transcriptional regulators of development often function through dispersed cis elements at endogenous target genes. While cis-elements are routinely studied in transfection and transgenic reporter assays, it is challenging to ascertain how they function in vivo. To address this problem in the context of the locus encoding the critical hematopoietic transcription factor Gata2, we engineered mice lacking a cluster of GATA motifs 2.8 kb upstream of the Gata2 transcriptional start site. We demonstrate that the -2.8 kb site confers maximal Gata2 expression in hematopoietic stem cells and specific hematopoietic progenitors. By contrast to our previous demonstration that a palindromic GATA motif at the neighboring -1.8 kb site maintains Gata2 repression in terminally differentiating erythroid cells, the -2.8 kb site was not required to initiate or maintain repression. These analyses reveal qualitatively distinct functions of 2 GATA motif-containing regions in vivo.

[1]  Henriette O'Geen,et al.  Discovering hematopoietic mechanisms through genome-wide analysis of GATA factor chromatin occupancy. , 2009, Molecular cell.

[2]  J. D. Engel,et al.  GATA Motifs Regulate Early Hematopoietic Lineage-Specific Expression of the Gata2 Gene , 2005, Molecular and Cellular Biology.

[3]  M. Groudine,et al.  Targeted deletion of 5'HS1 and 5'HS4 of the beta-globin locus control region reveals additive activity of the DNaseI hypersensitive sites. , 2001, Blood.

[4]  K. Akashi,et al.  [Myeloid lineage commitment from the hematopoietic stem cell]. , 2007, Arerugi = [Allergy].

[5]  Ernest Fraenkel,et al.  Insights into GATA-1-mediated gene activation versus repression via genome-wide chromatin occupancy analysis. , 2009, Molecular cell.

[6]  E. Bresnick,et al.  Dynamic GATA Factor Interplay at a Multicomponent Regulatory Region of the GATA-2 Locus* , 2005, Journal of Biological Chemistry.

[7]  Emery H. Bresnick,et al.  GATA Switches as Developmental Drivers* , 2010, The Journal of Biological Chemistry.

[8]  Berthold Göttgens,et al.  Gata2, Fli1, and Scl form a recursively wired gene-regulatory circuit during early hematopoietic development , 2007, Proceedings of the National Academy of Sciences.

[9]  Elaine Dzierzak,et al.  GATA-2 Plays Two Functionally Distinct Roles during the Ontogeny of Hematopoietic Stem Cells , 2004, The Journal of experimental medicine.

[10]  John Anastasi,et al.  Cooperative and antagonistic interplay between PU.1 and GATA-2 in the specification of myeloid cell fates. , 2002, Immunity.

[11]  D Orlic,et al.  Enforced expression of the GATA-2 transcription factor blocks normal hematopoiesis. , 1999, Blood.

[12]  S. Orkin,et al.  Transcription factor GATA-2 is required for proliferation/survival of early hematopoietic cells and mast cell formation, but not for erythroid and myeloid terminal differentiation. , 1997, Blood.

[13]  Jing Wu,et al.  GATA-1-dependent transcriptional repression of GATA-2 via disruption of positive autoregulation and domain-wide chromatin remodeling , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Kirby D. Johnson,et al.  Developmental control via GATA factor interplay at chromatin domains , 2005, Journal of cellular physiology.

[15]  G. Blobel,et al.  Distinct Functions of Dispersed GATA Factor Complexes at an Endogenous Gene Locus , 2006, Molecular and Cellular Biology.

[16]  Tariq Enver,et al.  Haploinsufficiency of GATA-2 perturbs adult hematopoietic stem-cell homeostasis. , 2005, Blood.

[17]  T. Schroeder,et al.  Improved prospective identification of megakaryocyte–erythrocyte progenitor cells , 2009, British journal of haematology.

[18]  Daniel G Tenen,et al.  The order of expression of transcription factors directs hierarchical specification of hematopoietic lineages. , 2006, Genes & development.

[19]  Stuart H. Orkin,et al.  An early haematopoietic defect in mice lacking the transcription factor GATA-2 , 1994, Nature.

[20]  S. Orkin,et al.  A Single cis Element Maintains Repression of the Key Developmental Regulator Gata2 , 2010, PLoS genetics.