Hematopoietic Signaling Mechanism Revealed from a Stem/Progenitor Cell Cistrome

Mol Cell. 2015 Jul 2;59(1):62-74. doi: 10.1016/j.molcel.2015.05.020. Epub 2015 Jun 11.

Abstract

Thousands of cis-elements in genomes are predicted to have vital functions. Although conservation, activity in surrogate assays, polymorphisms, and disease mutations provide functional clues, deletion from endogenous loci constitutes the gold-standard test. A GATA-2-binding, Gata2 intronic cis-element (+9.5) required for hematopoietic stem cell genesis in mice is mutated in a human immunodeficiency syndrome. Because +9.5 is the only cis-element known to mediate stem cell genesis, we devised a strategy to identify functionally comparable enhancers ("+9.5-like") genome-wide. Gene editing revealed +9.5-like activity to mediate GATA-2 occupancy, chromatin opening, and transcriptional activation. A +9.5-like element resided in Samd14, which encodes a protein of unknown function. Samd14 increased hematopoietic progenitor levels/activity and promoted signaling by a pathway vital for hematopoietic stem/progenitor cell regulation (stem cell factor/c-Kit), and c-Kit rescued Samd14 loss-of-function phenotypes. Thus, the hematopoietic stem/progenitor cell cistrome revealed a mediator of a signaling pathway that has broad importance for stem/progenitor cell biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Differentiation / genetics
  • Cell Line
  • GATA2 Transcription Factor / genetics*
  • Hematopoietic Stem Cells / metabolism*
  • Mice
  • Molecular Sequence Data
  • Proteins / genetics*
  • Proteins / metabolism
  • Proto-Oncogene Proteins c-kit / genetics*
  • RNA Interference
  • RNA, Small Interfering
  • Signal Transduction
  • Transcription, Genetic / genetics
  • Transcriptional Activation / genetics*

Substances

  • GATA2 Transcription Factor
  • Gata2 protein, mouse
  • Proteins
  • RNA, Small Interfering
  • Samd14 protein, mouse
  • Proto-Oncogene Proteins c-kit

Associated data

  • GEO/GSE68602