Evi-1 transforming and repressor activities are mediated by CtBP co-repressor proteins

Susan Palmer, Jean-Paul Brouillet, Anna Kilbey, Ruth Fulton, Mark Walker, Merlin Crossley, Chris Bartholomew

Research output: Contribution to journalArticle

Abstract

Ectopic production of the EVI1 transcriptional repressor zinc finger protein is seen in 4–6% of human acute myeloid leukemias. Overexpression also transforms Rat1 fibroblasts by an unknown mechanism, which is likely to be related to its role in leukemia and which depends upon its repressor activity. We show here that mutant murine Evi-1 proteins, lacking either the N-terminal zinc finger DNA binding domain or both DNA binding zinc finger clusters, function as dominant negative mutants by reverting the transformed phenotype of Evi-1 transformed Rat1 fibroblasts. The dominant negative activity of the non-DNA binding mutants suggests sequestration of transformation-specific cofactors and that recruitment of these cellular factors might mediate Evi-1 transforming activity.C-terminal bindingprotein (CtBP) co-repressor family proteins bind PLDLS-like motifs. We show that the murine Evi-1 repressor domain has two such sites, PFDLT (site a, amino acids 553–559) and PLDLS (site b, amino acids 584–590), which independently can bind CtBP family co-repressor proteins, with site b binding with higher affinity than site a. Functional analysis of specific CtBP binding mutants show site b is absolutely required to mediate both transformation of Rat1 fibroblasts and transcriptional repressor activity. This is the first demonstration that the biological activity of a mammalian cellular transcriptional repressor protein is mediated by CtBPs. Furthermore, it suggests that CtBP proteins are involved in the development of some acute leukemias and that blocking their ability to specifically interact with EVI1 might provide a target for the development of pharmacological therapeutic agents.

Original languageEnglish
Pages (from-to)25834-25840
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number28
DOIs
Publication statusPublished - 1 Jul 2001

Fingerprint

Co-Repressor Proteins
Zinc Fingers
Fibroblasts
Zinc
Leukemia
Repressor Proteins
Amino Acids
Proteins
DNA
Acute Myeloid Leukemia
Functional analysis
Bioactivity
Binding Sites
Pharmacology
Phenotype
Demonstrations
Therapeutics

Keywords

  • leukaemia
  • Evi1

Cite this

Palmer, Susan ; Brouillet, Jean-Paul ; Kilbey, Anna ; Fulton, Ruth ; Walker, Mark ; Crossley, Merlin ; Bartholomew, Chris. / Evi-1 transforming and repressor activities are mediated by CtBP co-repressor proteins. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 28. pp. 25834-25840.
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abstract = "Ectopic production of the EVI1 transcriptional repressor zinc finger protein is seen in 4–6{\%} of human acute myeloid leukemias. Overexpression also transforms Rat1 fibroblasts by an unknown mechanism, which is likely to be related to its role in leukemia and which depends upon its repressor activity. We show here that mutant murine Evi-1 proteins, lacking either the N-terminal zinc finger DNA binding domain or both DNA binding zinc finger clusters, function as dominant negative mutants by reverting the transformed phenotype of Evi-1 transformed Rat1 fibroblasts. The dominant negative activity of the non-DNA binding mutants suggests sequestration of transformation-specific cofactors and that recruitment of these cellular factors might mediate Evi-1 transforming activity.C-terminal bindingprotein (CtBP) co-repressor family proteins bind PLDLS-like motifs. We show that the murine Evi-1 repressor domain has two such sites, PFDLT (site a, amino acids 553–559) and PLDLS (site b, amino acids 584–590), which independently can bind CtBP family co-repressor proteins, with site b binding with higher affinity than site a. Functional analysis of specific CtBP binding mutants show site b is absolutely required to mediate both transformation of Rat1 fibroblasts and transcriptional repressor activity. This is the first demonstration that the biological activity of a mammalian cellular transcriptional repressor protein is mediated by CtBPs. Furthermore, it suggests that CtBP proteins are involved in the development of some acute leukemias and that blocking their ability to specifically interact with EVI1 might provide a target for the development of pharmacological therapeutic agents.",
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Palmer, S, Brouillet, J-P, Kilbey, A, Fulton, R, Walker, M, Crossley, M & Bartholomew, C 2001, 'Evi-1 transforming and repressor activities are mediated by CtBP co-repressor proteins', Journal of Biological Chemistry, vol. 276, no. 28, pp. 25834-25840. https://doi.org/10.1074/jbc.M102343200

Evi-1 transforming and repressor activities are mediated by CtBP co-repressor proteins. / Palmer, Susan; Brouillet, Jean-Paul; Kilbey, Anna; Fulton, Ruth; Walker, Mark; Crossley, Merlin; Bartholomew, Chris.

In: Journal of Biological Chemistry, Vol. 276, No. 28, 01.07.2001, p. 25834-25840.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Evi-1 transforming and repressor activities are mediated by CtBP co-repressor proteins

AU - Palmer, Susan

AU - Brouillet, Jean-Paul

AU - Kilbey, Anna

AU - Fulton, Ruth

AU - Walker, Mark

AU - Crossley, Merlin

AU - Bartholomew, Chris

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N2 - Ectopic production of the EVI1 transcriptional repressor zinc finger protein is seen in 4–6% of human acute myeloid leukemias. Overexpression also transforms Rat1 fibroblasts by an unknown mechanism, which is likely to be related to its role in leukemia and which depends upon its repressor activity. We show here that mutant murine Evi-1 proteins, lacking either the N-terminal zinc finger DNA binding domain or both DNA binding zinc finger clusters, function as dominant negative mutants by reverting the transformed phenotype of Evi-1 transformed Rat1 fibroblasts. The dominant negative activity of the non-DNA binding mutants suggests sequestration of transformation-specific cofactors and that recruitment of these cellular factors might mediate Evi-1 transforming activity.C-terminal bindingprotein (CtBP) co-repressor family proteins bind PLDLS-like motifs. We show that the murine Evi-1 repressor domain has two such sites, PFDLT (site a, amino acids 553–559) and PLDLS (site b, amino acids 584–590), which independently can bind CtBP family co-repressor proteins, with site b binding with higher affinity than site a. Functional analysis of specific CtBP binding mutants show site b is absolutely required to mediate both transformation of Rat1 fibroblasts and transcriptional repressor activity. This is the first demonstration that the biological activity of a mammalian cellular transcriptional repressor protein is mediated by CtBPs. Furthermore, it suggests that CtBP proteins are involved in the development of some acute leukemias and that blocking their ability to specifically interact with EVI1 might provide a target for the development of pharmacological therapeutic agents.

AB - Ectopic production of the EVI1 transcriptional repressor zinc finger protein is seen in 4–6% of human acute myeloid leukemias. Overexpression also transforms Rat1 fibroblasts by an unknown mechanism, which is likely to be related to its role in leukemia and which depends upon its repressor activity. We show here that mutant murine Evi-1 proteins, lacking either the N-terminal zinc finger DNA binding domain or both DNA binding zinc finger clusters, function as dominant negative mutants by reverting the transformed phenotype of Evi-1 transformed Rat1 fibroblasts. The dominant negative activity of the non-DNA binding mutants suggests sequestration of transformation-specific cofactors and that recruitment of these cellular factors might mediate Evi-1 transforming activity.C-terminal bindingprotein (CtBP) co-repressor family proteins bind PLDLS-like motifs. We show that the murine Evi-1 repressor domain has two such sites, PFDLT (site a, amino acids 553–559) and PLDLS (site b, amino acids 584–590), which independently can bind CtBP family co-repressor proteins, with site b binding with higher affinity than site a. Functional analysis of specific CtBP binding mutants show site b is absolutely required to mediate both transformation of Rat1 fibroblasts and transcriptional repressor activity. This is the first demonstration that the biological activity of a mammalian cellular transcriptional repressor protein is mediated by CtBPs. Furthermore, it suggests that CtBP proteins are involved in the development of some acute leukemias and that blocking their ability to specifically interact with EVI1 might provide a target for the development of pharmacological therapeutic agents.

KW - leukaemia

KW - Evi1

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DO - 10.1074/jbc.M102343200

M3 - Article

VL - 276

SP - 25834

EP - 25840

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

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ER -

Palmer S, Brouillet J-P, Kilbey A, Fulton R, Walker M, Crossley M et al. Evi-1 transforming and repressor activities are mediated by CtBP co-repressor proteins. Journal of Biological Chemistry. 2001 Jul 1;276(28):25834-25840. https://doi.org/10.1074/jbc.M102343200