title: Posterior-anterior gradient of zebrafish hes6 expression in the presomitic mesoderm is established by the combinatorial functions of the downstream enhancer and 3'UTR.
abstract: In vertebrates, the periodic formation of somites from the presomitic mesoderm (PSM) is driven by the molecular oscillator known as the segmentation clock. The hairy-related gene, hes6/her13.2, functions as a hub by dimerizing with other oscillators of the segmentation clock in zebrafish embryos. Although hes6 exhibits a posterior-anterior expression gradient in the posterior PSM with a peak at the tailbud, the detailed mechanisms underlying this unique expression pattern have not yet been clarified. By establishing several transgenic lines, we found that the transcriptional regulatory region downstream of hes6 in combination with the hes6 3'UTR recapitulates the endogenous gradient of hes6 mRNA expression. This downstream region, which we termed the PT enhancer, possessed several putative binding sites for the T-box and Ets transcription factors that were required for the regulatory activity. Indeed, the T-box transcription factor (Tbx16) and Ets transcription factor (Pea3) bound specifically to the putative binding sites and regulated the enhancer activity in zebrafish embryos. In addition, the 3'UTR of hes6 is required for recapitulation of the endogenous mRNA expression. Furthermore, the PT enhancer with the 3'UTR of hes6 responded to the inhibition of retinoic acid synthesis and fibroblast growth factor signaling in a manner similar to endogenous hes6. The results showed that transcriptional regulation by the T-box and Ets transcription factors, combined with the mRNA stability given by the 3'UTR, is responsible for the unique expression gradient of hes6 mRNA in the posterior PSM of zebrafish embryos.
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan. Electronic address: akawamur@mail.saitama-u.ac.jp.
authors.forename: Akinori
authors.initials: A
authors.lastname: Kawamura
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Ovara
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Ooka
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Kinoshita
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Hoshikawa
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Nakajo
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Yokota
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Yuuri
authors.initials: Y
authors.lastname: Fujino
authors.affiliation: Okazaki Institute for Integrative Bioscience & National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki, Aichi 444-8787, Japan.
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Higashijima
authors.affiliation: Okazaki Institute for Integrative Bioscience & National Institute for Basic Biology, National Institutes of Natural Sciences, Okazaki, Aichi 444-8787, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Takada
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Kyo
authors.initials: K
authors.lastname: Yamasu
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D019521:Body Patterning / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000378:metabolism; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008648:Mesoderm / Q000187:drug effects / Q000196:embryology* / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D019170:Somites / Q000187:drug effects / Q000196:embryology* / Q000378:metabolism; D013623:Tail / Q000196:embryology; D014212:Tretinoin / Q000494:pharmacology; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; D051792:Basic Helix-Loop-Helix Proteins; D005346:Fibroblast Growth Factors; D049452:Green Fluorescent Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D014212:Tretinoin; D029961:Zebrafish Proteins; C000608283:Hes6 protein, zebrafish; C485184:enhanced green fluorescent protein
keywords: 3′UTR; Expression gradient; Hairy-related gene; Somite; Transcriptional regulation; Zebrafish