Use of altered specificity mutants to probe a specific protein-protein interaction in differentiation: the GATA-1:FOG complex

Mol Cell. 1999 Feb;3(2):219-28. doi: 10.1016/s1097-2765(00)80312-3.

Abstract

GATA-1 and FOG (Friend of GATA-1) are each essential for erythroid and megakaryocyte development. FOG, a zinc finger protein, interacts with the amino (N) finger of GATA-1 and cooperates with GATA-1 to promote differentiation. To determine whether this interaction is critical for GATA-1 action, we selected GATA-1 mutants in yeast that fail to interact with FOG but retain normal DNA binding, as well a compensatory FOG mutant that restores interaction. These novel GATA-1 mutants do not promote erythroid differentiation of GATA-1- erythroid cells. Differentiation is rescued by the second-site FOG mutant. Thus, interaction of FOG with GATA-1 is essential for the function of GATA-1 in erythroid differentiation. These findings provide a paradigm for dissecting protein-protein associations involved in mammalian development.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cell Differentiation
  • DNA / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Erythroid Precursor Cells / metabolism
  • Erythroid-Specific DNA-Binding Factors
  • Erythropoiesis / physiology*
  • GATA1 Transcription Factor
  • Gene Expression Regulation
  • Genetic Complementation Test
  • Macromolecular Substances
  • Mice
  • Molecular Sequence Data
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae / genetics
  • Structure-Activity Relationship
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transfection
  • Zinc Fingers

Substances

  • Carrier Proteins
  • DNA-Binding Proteins
  • Erythroid-Specific DNA-Binding Factors
  • GATA1 Transcription Factor
  • Gata1 protein, mouse
  • Macromolecular Substances
  • Nuclear Proteins
  • Recombinant Fusion Proteins
  • Transcription Factors
  • Zfpm1 protein, mouse
  • DNA