Use of altered specificity mutants to probe a specific protein-protein interaction in differentiation: the GATA-1:FOG complex |
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Authors: | Crispino J D Lodish M B MacKay J P Orkin S H |
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Affiliation: | Division of Hematology-Oncology, Children's Hospital, Boston, Massachusetts 02115, USA. |
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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. |
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