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

John D. Crispino; Maya B. Lodish; Joel P. MacKay; Stuart H. Orkin

doi:10.1016/S1097-2765(00)80312-3

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

John D. Crispino, Maya B. Lodish, Joel P. MacKay, Stuart H. Orkin^*

^*Corresponding author for this work

Medicine, Hematology Oncology Division

Research output: Contribution to journal › Article › peer-review

216 Scopus citations

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.

Original language	English (US)
Pages (from-to)	219-228
Number of pages	10
Journal	Molecular cell
Volume	3
Issue number	2
DOIs	https://doi.org/10.1016/S1097-2765(00)80312-3
State	Published - Feb 1999

Funding

We thank the laboratory for discussions, Paul Mead for manuscript review, Alice Tsang for continued input, and Richard Goodman for the split two-hybrid system. Dennis Hom contributed to the subcloning of the split two-hybrid vectors. J. D. C. is supported by a fellowship from the Jane Coffin Childs Fund for Medical Research. S. H. O. is an Investigator of the Howard Hughes Medical Institute. Partial support was provided by a grant from the NIH.

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1016/S1097-2765(00)80312-3

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title = "Use of altered specificity mutants to probe a specific protein-protein interaction in differentiation: The GATA-1:FOG complex",

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.",

author = "Crispino, {John D.} and Lodish, {Maya B.} and MacKay, {Joel P.} and Orkin, {Stuart H.}",

note = "Funding Information: We thank the laboratory for discussions, Paul Mead for manuscript review, Alice Tsang for continued input, and Richard Goodman for the split two-hybrid system. Dennis Hom contributed to the subcloning of the split two-hybrid vectors. J. D. C. is supported by a fellowship from the Jane Coffin Childs Fund for Medical Research. S. H. O. is an Investigator of the Howard Hughes Medical Institute. Partial support was provided by a grant from the NIH.",

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doi = "10.1016/S1097-2765(00)80312-3",

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AU - MacKay, Joel P.

AU - Orkin, Stuart H.

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

Abstract

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