Sox3

A transcription factor for Cyp19 expression in the frog Rana rugosa

Yuki Oshima, Kiyoshi Naruse, Yoriko Nakamura, Masahisa Nakamura

    Research output: Contribution to journalArticle

    29 Citations (Scopus)

    Abstract

    Cyp19 is expressed at a high level in the gonad of the female tadpole of the frog Rana rugosa during sex determination. To identify sequence elements important for expression of Cyp19, we isolated a genomic clone (∼ 40 kbp) carrying R. rugosa Cyp19 and analyzed the nucleotide sequence of the 5′-flanking region to search for potential transcription factor binding sites. Sox (SRY-related HMG box) protein and Sf1 binding sites were found in the ovary-specific promoter region of Cyp19. Because Sox3 is located on the sex chromosome in R. rugosa, we conducted the luciferase reporter assay in Xenopus A6 cells using the promoter region. Sox3 drove the reporter gene in the cells, but Sf1 did not. When sequential deletion of the 2.7 kbp Cyp19-promoter region was undertaken, a fragment spanning nucleotides - 191 to + 48 was sufficient to drive the transcription of the reporter gene. In site-directed mutagenesis, the binding site at - 57 in the region was critical for Sox3 responsiveness. Sox3 lacking the HMG box had no ability to promote Cyp19 transcription. In addition, a chromatin immunoprecipitation (ChIP) assay showed that DNA fragments were enriched 8-fold, as determined by real-time PCR, when chromatin was immunoprecipitated with the anti-His antibody against His-tagged Sox3. The results, taken together, suggest that Sox3 activates Cyp19 transcription by its direct binding to the binding site of the Cyp19 promoter region. Sox3 appears to be a factor that directs indifferent gonads to develop into an ovary in R. rugosa.

    Original languageEnglish
    Pages (from-to)38-48
    Number of pages11
    JournalGene
    Volume445
    Issue number1-2
    DOIs
    Publication statusPublished - 2009 Sep 15

    Fingerprint

    Ranidae
    Genetic Promoter Regions
    Anura
    Transcription Factors
    Binding Sites
    Gonads
    Reporter Genes
    Ovary
    HMGB Proteins
    Sex Chromosomes
    Chromatin Immunoprecipitation
    5' Flanking Region
    Xenopus
    Site-Directed Mutagenesis
    Luciferases
    Protein Binding
    Chromatin
    Larva
    Real-Time Polymerase Chain Reaction
    Anti-Idiotypic Antibodies

    Keywords

    • Amphibians
    • ChIP assay
    • Cyp19
    • Promoter assay
    • Rana rugosa
    • Sex determination
    • Sox3

    ASJC Scopus subject areas

    • Genetics

    Cite this

    Oshima, Y., Naruse, K., Nakamura, Y., & Nakamura, M. (2009). Sox3: A transcription factor for Cyp19 expression in the frog Rana rugosa. Gene, 445(1-2), 38-48. https://doi.org/10.1016/j.gene.2009.05.011

    Sox3 : A transcription factor for Cyp19 expression in the frog Rana rugosa. / Oshima, Yuki; Naruse, Kiyoshi; Nakamura, Yoriko; Nakamura, Masahisa.

    In: Gene, Vol. 445, No. 1-2, 15.09.2009, p. 38-48.

    Research output: Contribution to journalArticle

    Oshima, Y, Naruse, K, Nakamura, Y & Nakamura, M 2009, 'Sox3: A transcription factor for Cyp19 expression in the frog Rana rugosa', Gene, vol. 445, no. 1-2, pp. 38-48. https://doi.org/10.1016/j.gene.2009.05.011
    Oshima, Yuki ; Naruse, Kiyoshi ; Nakamura, Yoriko ; Nakamura, Masahisa. / Sox3 : A transcription factor for Cyp19 expression in the frog Rana rugosa. In: Gene. 2009 ; Vol. 445, No. 1-2. pp. 38-48.
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