Characterization of the isoforms of MOVO zinc finger protein, a mouse homologue of Drosophila Ovo, as transcription factors

Sawako Unezaki, Mikio Nishizawa, Emiko Okuda-Ashitaka, Yasuo Masu, Masanori Mukai, Satoru Kobayashi, Kazunobu Sawamoto, Hideyuki Okano, Seiji Ito

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

We previously described two isoforms (MOVO-A and -B) of a novel zinc finger protein MOVO, a mouse homologue of Drosophila Ovo protein. Here, we isolated cDNA encoding the third isoform MOVO-C, which had a transactivation domain and zinc finger domain, but lacked an N-terminal potential repression domain that was present in MOVO-A. Three isoform mRNAs were expressed highly in mouse testis and also in the ovary at lower levels. The structural analyses of the isolated Movo gene and mRNAs demonstrated that three different Movo transcripts were differentially processed to generate three isoforms. Major mRNA species encoded MOVO-B with a zinc finger domain alone, and minor mRNA species encoded MOVO-A (potential repressor) and MOVO-C (potential activator). To assign MOVO to a transcriptional factor, we characterized DNA-binding and transactivation properties. Random oligonucleotide selection, electrophoretic mobility shift assay and footprinting indicated that MOVO bound to the sequence, 5′-G(G/C/T)GGGGG-3′. These motifs were found in the 5′-flanking regions of Movo and other testis-specific genes. Nuclear proteins binding to this motif were detected in mouse testis, and the expression of MOVO mRNA was restricted in spermatocytes. The luciferase assay demonstrated that MOVO-C activated Movo promoter and MOVO-A repressed it, but MOVO-B had no effects. Mutated MOVO-binding motifs in the Movo promoter reduced the luciferase activity. All the isoforms had no effects on SV40 promoter without MOVO-binding motifs. MOVO-A partially rescued oogenesis of a Drosophila ovo mutant. These results suggest that MOVO isoforms are transcription factors to regulate genes carrying the MOVO-binding motifs in the testis.

Original languageEnglish
Pages (from-to)47-58
Number of pages12
JournalGene
Volume336
Issue number1
DOIs
Publication statusPublished - 2004 Jul 7

Fingerprint

Zinc Fingers
Drosophila
Protein Isoforms
Transcription Factors
Testis
Messenger RNA
Proteins
Luciferases
Transcriptional Activation
Genes
RNA Isoforms
Oogenesis
Spermatocytes
5' Flanking Region
Electrophoretic Mobility Shift Assay
Nuclear Proteins
Protein Binding
Oligonucleotides
Ovary
Complementary DNA

Keywords

  • cDNA
  • DNA complementary to RNA
  • EF
  • elongation factor 1α
  • Oogenesis
  • otu
  • ovarian tumor
  • PAGE
  • PCR
  • polymerase chain reaction
  • RACE
  • rapid amplification of cDNA ends
  • reverse transcription
  • ribonuclease
  • RNA processing
  • RNase
  • RT
  • Spermatocyte
  • Testis

ASJC Scopus subject areas

  • Genetics

Cite this

Unezaki, S., Nishizawa, M., Okuda-Ashitaka, E., Masu, Y., Mukai, M., Kobayashi, S., ... Ito, S. (2004). Characterization of the isoforms of MOVO zinc finger protein, a mouse homologue of Drosophila Ovo, as transcription factors. Gene, 336(1), 47-58. https://doi.org/10.1016/j.gene.2004.03.013

Characterization of the isoforms of MOVO zinc finger protein, a mouse homologue of Drosophila Ovo, as transcription factors. / Unezaki, Sawako; Nishizawa, Mikio; Okuda-Ashitaka, Emiko; Masu, Yasuo; Mukai, Masanori; Kobayashi, Satoru; Sawamoto, Kazunobu; Okano, Hideyuki; Ito, Seiji.

In: Gene, Vol. 336, No. 1, 07.07.2004, p. 47-58.

Research output: Contribution to journalArticle

Unezaki, S, Nishizawa, M, Okuda-Ashitaka, E, Masu, Y, Mukai, M, Kobayashi, S, Sawamoto, K, Okano, H & Ito, S 2004, 'Characterization of the isoforms of MOVO zinc finger protein, a mouse homologue of Drosophila Ovo, as transcription factors', Gene, vol. 336, no. 1, pp. 47-58. https://doi.org/10.1016/j.gene.2004.03.013
Unezaki, Sawako ; Nishizawa, Mikio ; Okuda-Ashitaka, Emiko ; Masu, Yasuo ; Mukai, Masanori ; Kobayashi, Satoru ; Sawamoto, Kazunobu ; Okano, Hideyuki ; Ito, Seiji. / Characterization of the isoforms of MOVO zinc finger protein, a mouse homologue of Drosophila Ovo, as transcription factors. In: Gene. 2004 ; Vol. 336, No. 1. pp. 47-58.
@article{556cb4ed0777489faea8bc410a423e74,
title = "Characterization of the isoforms of MOVO zinc finger protein, a mouse homologue of Drosophila Ovo, as transcription factors",
abstract = "We previously described two isoforms (MOVO-A and -B) of a novel zinc finger protein MOVO, a mouse homologue of Drosophila Ovo protein. Here, we isolated cDNA encoding the third isoform MOVO-C, which had a transactivation domain and zinc finger domain, but lacked an N-terminal potential repression domain that was present in MOVO-A. Three isoform mRNAs were expressed highly in mouse testis and also in the ovary at lower levels. The structural analyses of the isolated Movo gene and mRNAs demonstrated that three different Movo transcripts were differentially processed to generate three isoforms. Major mRNA species encoded MOVO-B with a zinc finger domain alone, and minor mRNA species encoded MOVO-A (potential repressor) and MOVO-C (potential activator). To assign MOVO to a transcriptional factor, we characterized DNA-binding and transactivation properties. Random oligonucleotide selection, electrophoretic mobility shift assay and footprinting indicated that MOVO bound to the sequence, 5′-G(G/C/T)GGGGG-3′. These motifs were found in the 5′-flanking regions of Movo and other testis-specific genes. Nuclear proteins binding to this motif were detected in mouse testis, and the expression of MOVO mRNA was restricted in spermatocytes. The luciferase assay demonstrated that MOVO-C activated Movo promoter and MOVO-A repressed it, but MOVO-B had no effects. Mutated MOVO-binding motifs in the Movo promoter reduced the luciferase activity. All the isoforms had no effects on SV40 promoter without MOVO-binding motifs. MOVO-A partially rescued oogenesis of a Drosophila ovo mutant. These results suggest that MOVO isoforms are transcription factors to regulate genes carrying the MOVO-binding motifs in the testis.",
keywords = "cDNA, DNA complementary to RNA, EF, elongation factor 1α, Oogenesis, otu, ovarian tumor, PAGE, PCR, polymerase chain reaction, RACE, rapid amplification of cDNA ends, reverse transcription, ribonuclease, RNA processing, RNase, RT, Spermatocyte, Testis",
author = "Sawako Unezaki and Mikio Nishizawa and Emiko Okuda-Ashitaka and Yasuo Masu and Masanori Mukai and Satoru Kobayashi and Kazunobu Sawamoto and Hideyuki Okano and Seiji Ito",
year = "2004",
month = "7",
day = "7",
doi = "10.1016/j.gene.2004.03.013",
language = "English",
volume = "336",
pages = "47--58",
journal = "Gene",
issn = "0378-1119",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Characterization of the isoforms of MOVO zinc finger protein, a mouse homologue of Drosophila Ovo, as transcription factors

AU - Unezaki, Sawako

AU - Nishizawa, Mikio

AU - Okuda-Ashitaka, Emiko

AU - Masu, Yasuo

AU - Mukai, Masanori

AU - Kobayashi, Satoru

AU - Sawamoto, Kazunobu

AU - Okano, Hideyuki

AU - Ito, Seiji

PY - 2004/7/7

Y1 - 2004/7/7

N2 - We previously described two isoforms (MOVO-A and -B) of a novel zinc finger protein MOVO, a mouse homologue of Drosophila Ovo protein. Here, we isolated cDNA encoding the third isoform MOVO-C, which had a transactivation domain and zinc finger domain, but lacked an N-terminal potential repression domain that was present in MOVO-A. Three isoform mRNAs were expressed highly in mouse testis and also in the ovary at lower levels. The structural analyses of the isolated Movo gene and mRNAs demonstrated that three different Movo transcripts were differentially processed to generate three isoforms. Major mRNA species encoded MOVO-B with a zinc finger domain alone, and minor mRNA species encoded MOVO-A (potential repressor) and MOVO-C (potential activator). To assign MOVO to a transcriptional factor, we characterized DNA-binding and transactivation properties. Random oligonucleotide selection, electrophoretic mobility shift assay and footprinting indicated that MOVO bound to the sequence, 5′-G(G/C/T)GGGGG-3′. These motifs were found in the 5′-flanking regions of Movo and other testis-specific genes. Nuclear proteins binding to this motif were detected in mouse testis, and the expression of MOVO mRNA was restricted in spermatocytes. The luciferase assay demonstrated that MOVO-C activated Movo promoter and MOVO-A repressed it, but MOVO-B had no effects. Mutated MOVO-binding motifs in the Movo promoter reduced the luciferase activity. All the isoforms had no effects on SV40 promoter without MOVO-binding motifs. MOVO-A partially rescued oogenesis of a Drosophila ovo mutant. These results suggest that MOVO isoforms are transcription factors to regulate genes carrying the MOVO-binding motifs in the testis.

AB - We previously described two isoforms (MOVO-A and -B) of a novel zinc finger protein MOVO, a mouse homologue of Drosophila Ovo protein. Here, we isolated cDNA encoding the third isoform MOVO-C, which had a transactivation domain and zinc finger domain, but lacked an N-terminal potential repression domain that was present in MOVO-A. Three isoform mRNAs were expressed highly in mouse testis and also in the ovary at lower levels. The structural analyses of the isolated Movo gene and mRNAs demonstrated that three different Movo transcripts were differentially processed to generate three isoforms. Major mRNA species encoded MOVO-B with a zinc finger domain alone, and minor mRNA species encoded MOVO-A (potential repressor) and MOVO-C (potential activator). To assign MOVO to a transcriptional factor, we characterized DNA-binding and transactivation properties. Random oligonucleotide selection, electrophoretic mobility shift assay and footprinting indicated that MOVO bound to the sequence, 5′-G(G/C/T)GGGGG-3′. These motifs were found in the 5′-flanking regions of Movo and other testis-specific genes. Nuclear proteins binding to this motif were detected in mouse testis, and the expression of MOVO mRNA was restricted in spermatocytes. The luciferase assay demonstrated that MOVO-C activated Movo promoter and MOVO-A repressed it, but MOVO-B had no effects. Mutated MOVO-binding motifs in the Movo promoter reduced the luciferase activity. All the isoforms had no effects on SV40 promoter without MOVO-binding motifs. MOVO-A partially rescued oogenesis of a Drosophila ovo mutant. These results suggest that MOVO isoforms are transcription factors to regulate genes carrying the MOVO-binding motifs in the testis.

KW - cDNA

KW - DNA complementary to RNA

KW - EF

KW - elongation factor 1α

KW - Oogenesis

KW - otu

KW - ovarian tumor

KW - PAGE

KW - PCR

KW - polymerase chain reaction

KW - RACE

KW - rapid amplification of cDNA ends

KW - reverse transcription

KW - ribonuclease

KW - RNA processing

KW - RNase

KW - RT

KW - Spermatocyte

KW - Testis

UR - http://www.scopus.com/inward/record.url?scp=3242742164&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3242742164&partnerID=8YFLogxK

U2 - 10.1016/j.gene.2004.03.013

DO - 10.1016/j.gene.2004.03.013

M3 - Article

VL - 336

SP - 47

EP - 58

JO - Gene

JF - Gene

SN - 0378-1119

IS - 1

ER -