Transcription control of a gene for Drosophila transcription factor, DREF by DRE and cis-elements conserved between Drosophila melanogaster and virilis

Eunjeong Kwon, Hirokazu Seto, Fumiko Hirose, Nobuko Oshima, Yasuhiko Takahashi, Yasuyoshi Nishida, Masamitsu Yamaguchi

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A DNA replication-related element (DRE)-binding factor (DREF) has been revealed to be an important transcription factor for activating promoters of cell proliferation and differentiation related genes. The amino acid sequences of DREF are conserved in evolutionary separate Drosophila species, Drosophila melanogaster (Dm) and Drosophila virilis (Dv) in three regions. In the present study, evidence was obtained that there are several highly conserved regions in the 5′ flanking region between the DmDREF and DvDREF genes. Band mobility shift assays using oligonucleotides corresponding to these conserved regions revealed that specific trans-acting factors can bind to at least three regions -554 to -543 (5′-TTTGTTCTTGCG), -81 to -70 (5′-GCCCACGTGGCT) and +225 to +234 (5′-GCAATCAGTG). Using a transient luciferase expression assay, we demonstrated that the region -554 to -543 functions as a negative regulatory element for DmDREF promoter activity, while the regions -77 to -70 (5′-ACGTGGCT) and +225 to +236 (5′-GCAATCAGTGTT) function as positive regulatory elements. In previous studies, we observed that expression of the homeodomain protein Zerknüllt (Zen) represses PCNA gene transcription, by reducing the DNA binding activity of DREF. Here we show Zen downregulates DREF gene promoter activity through action on the region between +241 and +254 (5′-AGAATACTCAACA). In addition, the DmDREF promoter contains five DREs. Using a double stranded RNA-mediated interference method, we generated evidence that expression of DmDREF could be auto-regulated by DREF through the third DRE located at +211 to +218. In living flies we obtained results consistent with those obtained in vitro and in cultured cells. The study thus indicates that DmDREF is effectively regulated via highly conserved regions between the DmDREF and DvDREF promoters, suggesting the existence of common regulatory factors, and that DmDREF can be positively regulated by itself via the third DRE located in its most highly conserved region.

Original languageEnglish
Pages (from-to)101-116
Number of pages16
JournalGene
Volume309
Issue number2
DOIs
Publication statusPublished - 08-05-2003

Fingerprint

Drosophila melanogaster
DNA Replication
Drosophila
Transcription Factors
Genes
Activating Transcription Factors
Homeodomain Proteins
Trans-Activators
Double-Stranded RNA
5' Flanking Region
Proliferating Cell Nuclear Antigen
Electrophoretic Mobility Shift Assay
RNA Interference
Luciferases
Oligonucleotides
Diptera
Cell Differentiation
Amino Acid Sequence
Cultured Cells
Down-Regulation

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Kwon, Eunjeong ; Seto, Hirokazu ; Hirose, Fumiko ; Oshima, Nobuko ; Takahashi, Yasuhiko ; Nishida, Yasuyoshi ; Yamaguchi, Masamitsu. / Transcription control of a gene for Drosophila transcription factor, DREF by DRE and cis-elements conserved between Drosophila melanogaster and virilis. In: Gene. 2003 ; Vol. 309, No. 2. pp. 101-116.
@article{d3a9aba6f3454827a64ba473a3af7ba1,
title = "Transcription control of a gene for Drosophila transcription factor, DREF by DRE and cis-elements conserved between Drosophila melanogaster and virilis",
abstract = "A DNA replication-related element (DRE)-binding factor (DREF) has been revealed to be an important transcription factor for activating promoters of cell proliferation and differentiation related genes. The amino acid sequences of DREF are conserved in evolutionary separate Drosophila species, Drosophila melanogaster (Dm) and Drosophila virilis (Dv) in three regions. In the present study, evidence was obtained that there are several highly conserved regions in the 5′ flanking region between the DmDREF and DvDREF genes. Band mobility shift assays using oligonucleotides corresponding to these conserved regions revealed that specific trans-acting factors can bind to at least three regions -554 to -543 (5′-TTTGTTCTTGCG), -81 to -70 (5′-GCCCACGTGGCT) and +225 to +234 (5′-GCAATCAGTG). Using a transient luciferase expression assay, we demonstrated that the region -554 to -543 functions as a negative regulatory element for DmDREF promoter activity, while the regions -77 to -70 (5′-ACGTGGCT) and +225 to +236 (5′-GCAATCAGTGTT) function as positive regulatory elements. In previous studies, we observed that expression of the homeodomain protein Zerkn{\"u}llt (Zen) represses PCNA gene transcription, by reducing the DNA binding activity of DREF. Here we show Zen downregulates DREF gene promoter activity through action on the region between +241 and +254 (5′-AGAATACTCAACA). In addition, the DmDREF promoter contains five DREs. Using a double stranded RNA-mediated interference method, we generated evidence that expression of DmDREF could be auto-regulated by DREF through the third DRE located at +211 to +218. In living flies we obtained results consistent with those obtained in vitro and in cultured cells. The study thus indicates that DmDREF is effectively regulated via highly conserved regions between the DmDREF and DvDREF promoters, suggesting the existence of common regulatory factors, and that DmDREF can be positively regulated by itself via the third DRE located in its most highly conserved region.",
author = "Eunjeong Kwon and Hirokazu Seto and Fumiko Hirose and Nobuko Oshima and Yasuhiko Takahashi and Yasuyoshi Nishida and Masamitsu Yamaguchi",
year = "2003",
month = "5",
day = "8",
doi = "10.1016/S0378-1119(03)00493-1",
language = "English",
volume = "309",
pages = "101--116",
journal = "Gene",
issn = "0378-1119",
publisher = "Elsevier",
number = "2",

}

Transcription control of a gene for Drosophila transcription factor, DREF by DRE and cis-elements conserved between Drosophila melanogaster and virilis. / Kwon, Eunjeong; Seto, Hirokazu; Hirose, Fumiko; Oshima, Nobuko; Takahashi, Yasuhiko; Nishida, Yasuyoshi; Yamaguchi, Masamitsu.

In: Gene, Vol. 309, No. 2, 08.05.2003, p. 101-116.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Transcription control of a gene for Drosophila transcription factor, DREF by DRE and cis-elements conserved between Drosophila melanogaster and virilis

AU - Kwon, Eunjeong

AU - Seto, Hirokazu

AU - Hirose, Fumiko

AU - Oshima, Nobuko

AU - Takahashi, Yasuhiko

AU - Nishida, Yasuyoshi

AU - Yamaguchi, Masamitsu

PY - 2003/5/8

Y1 - 2003/5/8

N2 - A DNA replication-related element (DRE)-binding factor (DREF) has been revealed to be an important transcription factor for activating promoters of cell proliferation and differentiation related genes. The amino acid sequences of DREF are conserved in evolutionary separate Drosophila species, Drosophila melanogaster (Dm) and Drosophila virilis (Dv) in three regions. In the present study, evidence was obtained that there are several highly conserved regions in the 5′ flanking region between the DmDREF and DvDREF genes. Band mobility shift assays using oligonucleotides corresponding to these conserved regions revealed that specific trans-acting factors can bind to at least three regions -554 to -543 (5′-TTTGTTCTTGCG), -81 to -70 (5′-GCCCACGTGGCT) and +225 to +234 (5′-GCAATCAGTG). Using a transient luciferase expression assay, we demonstrated that the region -554 to -543 functions as a negative regulatory element for DmDREF promoter activity, while the regions -77 to -70 (5′-ACGTGGCT) and +225 to +236 (5′-GCAATCAGTGTT) function as positive regulatory elements. In previous studies, we observed that expression of the homeodomain protein Zerknüllt (Zen) represses PCNA gene transcription, by reducing the DNA binding activity of DREF. Here we show Zen downregulates DREF gene promoter activity through action on the region between +241 and +254 (5′-AGAATACTCAACA). In addition, the DmDREF promoter contains five DREs. Using a double stranded RNA-mediated interference method, we generated evidence that expression of DmDREF could be auto-regulated by DREF through the third DRE located at +211 to +218. In living flies we obtained results consistent with those obtained in vitro and in cultured cells. The study thus indicates that DmDREF is effectively regulated via highly conserved regions between the DmDREF and DvDREF promoters, suggesting the existence of common regulatory factors, and that DmDREF can be positively regulated by itself via the third DRE located in its most highly conserved region.

AB - A DNA replication-related element (DRE)-binding factor (DREF) has been revealed to be an important transcription factor for activating promoters of cell proliferation and differentiation related genes. The amino acid sequences of DREF are conserved in evolutionary separate Drosophila species, Drosophila melanogaster (Dm) and Drosophila virilis (Dv) in three regions. In the present study, evidence was obtained that there are several highly conserved regions in the 5′ flanking region between the DmDREF and DvDREF genes. Band mobility shift assays using oligonucleotides corresponding to these conserved regions revealed that specific trans-acting factors can bind to at least three regions -554 to -543 (5′-TTTGTTCTTGCG), -81 to -70 (5′-GCCCACGTGGCT) and +225 to +234 (5′-GCAATCAGTG). Using a transient luciferase expression assay, we demonstrated that the region -554 to -543 functions as a negative regulatory element for DmDREF promoter activity, while the regions -77 to -70 (5′-ACGTGGCT) and +225 to +236 (5′-GCAATCAGTGTT) function as positive regulatory elements. In previous studies, we observed that expression of the homeodomain protein Zerknüllt (Zen) represses PCNA gene transcription, by reducing the DNA binding activity of DREF. Here we show Zen downregulates DREF gene promoter activity through action on the region between +241 and +254 (5′-AGAATACTCAACA). In addition, the DmDREF promoter contains five DREs. Using a double stranded RNA-mediated interference method, we generated evidence that expression of DmDREF could be auto-regulated by DREF through the third DRE located at +211 to +218. In living flies we obtained results consistent with those obtained in vitro and in cultured cells. The study thus indicates that DmDREF is effectively regulated via highly conserved regions between the DmDREF and DvDREF promoters, suggesting the existence of common regulatory factors, and that DmDREF can be positively regulated by itself via the third DRE located in its most highly conserved region.

UR - http://www.scopus.com/inward/record.url?scp=0038324365&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0038324365&partnerID=8YFLogxK

U2 - 10.1016/S0378-1119(03)00493-1

DO - 10.1016/S0378-1119(03)00493-1

M3 - Article

C2 - 12758126

AN - SCOPUS:0038324365

VL - 309

SP - 101

EP - 116

JO - Gene

JF - Gene

SN - 0378-1119

IS - 2

ER -