Mechanism governing a stem cell-generating cis-regulatory element

Rajendran Sanalkumar, Kirby D. Johnson, Xin Gao, Meghan E. Boyer, Yuan I. Chang, Kyle J. Hewitt, Jing Zhang, Emery H. Bresnick

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

The unremitting demand to replenish differentiated cells in tissues requires efficient mechanisms to generate and regulate stem and progenitor cells. Although master regulatory transcription factors, including GATA binding protein-2 (GATA-2), have crucial roles in these mechanisms, how such factors are controlled in developmentally dynamic systems is poorly understood. Previously, we described five dispersed Gata2 locus sequences, termed the -77, -3.9, -2.8, -1.8, and +9.5 GATA switch sites, which contain evolutionarily conserved GATA motifs occupied by GATA-2 and GATA-1 in hematopoietic precursors and erythroid cells, respectively. Despite common attributes of transcriptional enhancers, targeted deletions of the -2.8, -1.8, and +9.5 sites revealed distinct and unpredictable contributions to Gata2 expression and hematopoiesis. Herein, we describe the targeted deletion of the -3.9 site and mechanistically compare the -3.9 sitewith other GATA switch sites. The -3.9-/- mice were viable and exhibited normal Gata2 expression and steady-state hematopoiesis in the embryo and adult. We established a Gata2 repression/reactivation assay, which revealed unique +9.5 site activity to mediate GATA factor-dependent chromatin structural transitions. Loss-of-function analyses provided evidence for a mechanism in which a mediator of long-range transcriptional control [LIM domain binding 1 (LDB1)] and a chromatin remodeler [Brahma related gene 1 (BRG1)] synergize through the +9.5 site, conferring expression of GATA-2, which is known to promote the genesis and survival of hematopoietic stem cells.

Original languageEnglish (US)
Pages (from-to)E1091-E1100
JournalProceedings of the National Academy of Sciences of the United States of America
Volume111
Issue number12
DOIs
StatePublished - Mar 25 2014

    Fingerprint

Keywords

  • Cis element
  • HSCs

ASJC Scopus subject areas

  • General

Cite this

Sanalkumar, R., Johnson, K. D., Gao, X., Boyer, M. E., Chang, Y. I., Hewitt, K. J., Zhang, J., & Bresnick, E. H. (2014). Mechanism governing a stem cell-generating cis-regulatory element. Proceedings of the National Academy of Sciences of the United States of America, 111(12), E1091-E1100. https://doi.org/10.1073/pnas.1400065111