Drosophila Mi-2 negatively regulates dDREF by inhibiting its DNA-binding activity

Fumiko Hirose, Nobuko Oshima, Eun Jeong Kwon, Hideki Yoshida, Masamitsu Yamaguchi

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Drosophila melanogaster DNA replication-related element (DRE) factor (dDREF) is a transcriptional regulatory factor required for the expression of genes carrying the 5′-TATCGATA DRE. dDREF has been reported to bind to a sequence in the chromatin boundary element, and thus, dDREF may play a part in regulating insulator activity. To generate further insights into dDREF function, we carried out a Saccharomyces cerevisiae two-hybrid screening with DREF polypeptide as bait and identified Mi-2 as a DREF-interacting protein. Biochemical analyses revealed that the C-terminal region of Drosophila Mi-2 (dMi-2) specifically binds to the DNA-binding domain of dDREF. Electrophoretic mobility shift assays showed that dMi-2 thereby inhibits the DNA-binding activity of dDREF. Ectopic expression of dDREF and dMi-2 in eye imaginal discs resulted in severe and mild rough-eye phenotypes, respectively, whereas flies simultaneously expressing both proteins exhibited almost-normal eye phenotypes. Half-dose reduction of the dMi-2 gene enhanced the DREF-induced rough-eye phenotype. Immunostaining of polytene chromosomes of salivary glands showed that dDREF and dMi-2 bind in mutually exclusive ways. These lines of evidence define a novel function of dMi-2 in the negative regulation of dDREF by its DNA-binding activity. Finally, we postulated that dDREF and dMi-2 may demonstrate reciprocal regulation of their functions.

Original languageEnglish
Pages (from-to)5182-5193
Number of pages12
JournalMolecular and Cellular Biology
Volume22
Issue number14
DOIs
Publication statusPublished - 09-07-2002

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Drosophila
DNA
Phenotype
Insulator Elements
Polytene Chromosomes
Imaginal Discs
Electrophoretic Mobility Shift Assay
Salivary Glands
Drosophila melanogaster
DNA Replication
Diptera
Chromatin
Saccharomyces cerevisiae
Proteins
Gene Expression
Peptides
Genes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Hirose, Fumiko ; Oshima, Nobuko ; Kwon, Eun Jeong ; Yoshida, Hideki ; Yamaguchi, Masamitsu. / Drosophila Mi-2 negatively regulates dDREF by inhibiting its DNA-binding activity. In: Molecular and Cellular Biology. 2002 ; Vol. 22, No. 14. pp. 5182-5193.
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Drosophila Mi-2 negatively regulates dDREF by inhibiting its DNA-binding activity. / Hirose, Fumiko; Oshima, Nobuko; Kwon, Eun Jeong; Yoshida, Hideki; Yamaguchi, Masamitsu.

In: Molecular and Cellular Biology, Vol. 22, No. 14, 09.07.2002, p. 5182-5193.

Research output: Contribution to journalArticle

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