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Nucleic Acids Res
2008 Jul 01;3612:3905-15. doi: 10.1093/nar/gkn291.
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Transcriptional regulation of the Drosophila moira and osa genes by the DREF pathway.
Nakamura K
,
Ida H
,
Yamaguchi M
.
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The DNA replication-related element binding factor (DREF) plays an important role in regulation of cell proliferation in Drosophila, binding to DRE and activating transcription of genes carrying this element in their promoter regions. Overexpression of DREF in eye imaginal discs induces a rough eye phenotype in adults, which can be suppressed by half dose reduction of the osa or moira (mor) genes encoding subunits of the BRM complex. This ATP-dependent chromatin remodeling complex is known to control gene expression and the cell cycle. In the 5'' flanking regions of the osa and mor genes, DRE and DRE-like sequences exist which contribute to their promoter activities. Expression levels and promoter activities of osa and mor are decreased in DREF knockdown cells and our results in vitro and in cultured cells indicate that transcription of osa and mor is regulated by the DRE/DREF regulatory pathway. In addition, mRNA levels of other BRM complex subunits and a target gene, string/cdc25, were found to be decreased by knockdown of DREF. These results indicate that DREF is involved in regulation of the BRM complex and thereby the cell cycle.
Figure 1. osa and mor genes genetically interact with DREF. Scanning electron micrographs of adult eyes. DREF overexpressing flies (GMR-GAL4/GMR-GAL4; UAS-DREF/UAS-DREF) were crossed with osa or mor mutant flies and developed at 28°C. (A, B) GMR-GAL4/+; UAS-DREF/+; +/+. (C, D) GMR-GAL4/+; UAS-DREF/+; osa2/+. (E, F) GMR-GAL4/+; UAS-DREF/+; P{PZ}osa00090/+. (G, H) GMR-GAL4/+; UAS-DREF/+; mor1/+. (A, C, E, G) Scale bars are for 50 μm. (B, D, F, H) Scale bars are for 12.5 μm.
Figure 2. DRE and DRE-like sequences in the 5′ flanking regions of the osa and mor genes. Transcription initiation site is numbered as +1. (A) A schematic representation of DREs in the osa gene. (B) A schematic representation of DREs in the mor gene.
Figure 3. Effects of mutations in DRE sites in the 5′ flanking region of the osa and mor genes on their promoter activities. osa or mor promoter–luciferase fusion plasmids were transfected into S2 cells. Luciferase activities are expressed relative to wild-type osa (A) and mor (B) promoters. Mean values with standard deviations from three independent transfections are shown. The white box, wild-type DRE; the black box, mutant DRE.
Figure 4. DREFdsRNA treatment affects mRNA levels and promoter activities of osa and mor. (A) cDNAs were prepared from total RNA isolated from dsRNA-treated S2 cells and levels of DREF, osa and mor mRNAs were measured by quantitative RT-PCR. Fold differences against the amplification with no treatment are shown with standard deviations from three independent dsRNA treatments. β-tubulin was used as a negative control. (B) Cell extracts were analyzed by western blotting with antiDREF antibodies. (C) Promoter activities of osa and mor in dsRNA-treated cells. Luciferase activities relative to that of wild-type promoter in non-dsRNA-treated cells are shown with standard deviations from five independent transfections.
Figure 5. Complex formation between osa DREs and Kc cell nuclear extracts. 32P-labeled double-stranded oligonucleotides osaDRE1 (A) and osaDRE2 (B) were incubated with Kc cell nuclear extracts in the presence of the indicated competitor oligonucleotides or antiDREF monoclonal antibodies. The amounts of competitors were 25-, 50- or 100-fold molar ratios for the osaDRE1 probe (A) and 100- or 400-fold molar ratios for the osaDRE2 probe (B). Mock, normal mouse IgG; Mab1, antiDREF monoclonal antibody 1; Mab4, antiDREF monoclonal antibody 4.
Figure 6. Complex formation between mor DREs and Kc cell nuclear extracts. 32P-labeled double-stranded oligonucleotides morDRE1 (A), morDRE2 (B) and morDRE3 (C) were incubated with Kc cell nuclear extracts in the presence of the indicated competitor oligonucleotides or antiDREF monoclonal antibodies. The amounts of competitors were 25-, 50- or 100-fold molar ratios for the morDRE1 probe (A) and 100- or 400-fold molar ratios for the morDRE2 (B) and morDRE3 probes (C). Mock, normal mouse IgG; Mab1, antiDREF monoclonal antibody 1; Mab4, antiDREF monoclonal antibody 4.
Figure 7. Binding of DREF to the DRE-containing genomic regions of the osa and mor genes. Crosslinked chromatin of S2 cells was immunoprecipitated with either antiDREF IgG or control rabbit IgG. The genomic regions containing osaDRE1, osaDRE2, morDRE1, 2 or morDRE3 were amplified by real-time PCR and compared with the amplification from the immunoprecipitates with the control IgG.
Figure 8. Effects of DREF knockdown on mRNA levels of other BRM complex components. (A) Total RNA was isolated from S2 cells treated with dsRNA and cDNA was prepared. mRNA levels were determined by quantitative RT-PCR. Fold differences versus no dsRNA treatment are shown as mean values with standard deviations from three independent dsRNA transfections. (B) DRE-containing regions were amplified from immunoprecipitates with antiDREF IgG. (C) mRNA level of stg in DREFdsRNA-treated cells.
Bouazoune,
ATP-dependent chromatin remodeling complexes in Drosophila.
2006, Pubmed
Bouazoune,
ATP-dependent chromatin remodeling complexes in Drosophila.
2006,
Pubmed
Brumby,
Drosophila cyclin E interacts with components of the Brahma complex.
2002,
Pubmed
,
Echinobase
Collins,
Osa-containing Brahma chromatin remodeling complexes are required for the repression of wingless target genes.
2000,
Pubmed
Collins,
Osa associates with the Brahma chromatin remodeling complex and promotes the activation of some target genes.
1999,
Pubmed
Crosby,
The trithorax group gene moira encodes a brahma-associated putative chromatin-remodeling factor in Drosophila melanogaster.
1999,
Pubmed
,
Echinobase
Cross,
Cell culture of individual Drosophila embryos. I. Development of wild-type cultures.
1978,
Pubmed
Edgar,
Genetic control of cell division patterns in the Drosophila embryo.
1989,
Pubmed
Elfring,
Identification and characterization of Drosophila relatives of the yeast transcriptional activator SNF2/SWI2.
1994,
Pubmed
Hayashi,
Drosophila distal-less negatively regulates dDREF by inhibiting its DNA binding activity.
2006,
Pubmed
Heitzler,
Enhancer-promoter communication mediated by Chip during Pannier-driven proneural patterning is regulated by Osa.
2003,
Pubmed
Hirose,
Ectopic expression of DREF induces DNA synthesis, apoptosis, and unusual morphogenesis in the Drosophila eye imaginal disc: possible interaction with Polycomb and trithorax group proteins.
2001,
Pubmed
,
Echinobase
Hirose,
Drosophila Mi-2 negatively regulates dDREF by inhibiting its DNA-binding activity.
2002,
Pubmed
Hirose,
Novel 8-base pair sequence (Drosophila DNA replication-related element) and specific binding factor involved in the expression of Drosophila genes for DNA polymerase alpha and proliferating cell nuclear antigen.
1993,
Pubmed
Hirose,
Isolation and characterization of cDNA for DREF, a promoter-activating factor for Drosophila DNA replication-related genes.
1996,
Pubmed
Hochheimer,
TRF2 associates with DREF and directs promoter-selective gene expression in Drosophila.
2002,
Pubmed
Hyun,
DREF is required for efficient growth and cell cycle progression in Drosophila imaginal discs.
2005,
Pubmed
Ida,
Identification of the Drosophila eIF4A gene as a target of the DREF transcription factor.
2007,
Pubmed
Jasper,
A genomic switch at the transition from cell proliferation to terminal differentiation in the Drosophila eye.
2002,
Pubmed
Kennison,
Dosage-dependent modifiers of polycomb and antennapedia mutations in Drosophila.
1988,
Pubmed
Kwon,
Transcriptional regulation of the Drosophila raf proto-oncogene by Drosophila STAT during development and in immune response.
2000,
Pubmed
Luo,
Rb interacts with histone deacetylase to repress transcription.
1998,
Pubmed
Matsukage,
The DRE/DREF transcriptional regulatory system: a master key for cell proliferation.
2008,
Pubmed
Mohrmann,
Differential targeting of two distinct SWI/SNF-related Drosophila chromatin-remodeling complexes.
2004,
Pubmed
Mohrmann,
Composition and functional specificity of SWI2/SNF2 class chromatin remodeling complexes.
2005,
Pubmed
Moshkin,
Functional differentiation of SWI/SNF remodelers in transcription and cell cycle control.
2007,
Pubmed
Ohler,
Computational analysis of core promoters in the Drosophila genome.
2002,
Pubmed
Okudaira,
Transcriptional regulation of the Drosophila orc2 gene by the DREF pathway.
2005,
Pubmed
Papoulas,
The Drosophila trithorax group proteins BRM, ASH1 and ASH2 are subunits of distinct protein complexes.
1998,
Pubmed
,
Echinobase
Phuong Thao,
Identification of the Drosophila skpA gene as a novel target of the transcription factor DREF.
2006,
Pubmed
Rabenstein,
TATA box-binding protein (TBP)-related factor 2 (TRF2), a third member of the TBP family.
1999,
Pubmed
Sawado,
The DNA replication-related element (DRE)/DRE-binding factor system is a transcriptional regulator of the Drosophila E2F gene.
1998,
Pubmed
Sohn,
SRG3 interacts directly with the major components of the SWI/SNF chromatin remodeling complex and protects them from proteasomal degradation.
2007,
Pubmed
,
Echinobase
Staehling-Hampton,
A genetic screen for modifiers of E2F in Drosophila melanogaster.
1999,
Pubmed
,
Echinobase
Takahashi,
DNA replication-related elements cooperate to enhance promoter activity of the drosophila DNA polymerase alpha 73-kDa subunit gene.
1996,
Pubmed
Takahashi,
Identification of three conserved regions in the DREF transcription factors from Drosophila melanogaster and Drosophila virilis.
1999,
Pubmed
Treisman,
eyelid antagonizes wingless signaling during Drosophila development and has homology to the Bright family of DNA-binding proteins.
1997,
Pubmed
Tsuchiya,
Transcriptional regulation of the Drosophila rfc1 gene by the DRE-DREF pathway.
2007,
Pubmed
Yamaguchi,
A nucleotide sequence essential for the function of DRE, a common promoter element for Drosophila DNa replication-related genes.
1995,
Pubmed
Yamaguchi,
Ectopic expression of human p53 inhibits entry into S phase and induces apoptosis in the Drosophila eye imaginal disc.
1999,
Pubmed
Yoshida,
DREF is required for EGFR signalling during Drosophila wing vein development.
2004,
Pubmed