Identification of ZAG1, a Novel Protein Expressed in Mouse Preimplantation, and Its Putative Roles in Zygotic Genome Activation

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  • MATSUOKA Toshiki
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University
  • SATO Manabu
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University
  • TOKORO Mikiko
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University
  • SHIN Seung-Wook
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University
  • UENOYAMA Atsuto
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University
  • ITO Kazunari
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University
  • HITOMI Syuji
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University
  • AMANO Tomoko
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University
  • ANZAI Masayuki
    Institute of Advanced technology, Kinki University
  • KATO Hiromi
    Institute of Advanced technology, Kinki University
  • MITANI Tasuku
    Institute of Advanced technology, Kinki University
  • SAEKI Kazuhiro
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University Institute of Advanced technology, Kinki University
  • HOSOI Yoshihiko
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University Institute of Advanced technology, Kinki University
  • IRITANI Akira
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University Institute of Advanced technology, Kinki University
  • MATSUMOTO Kazuya
    Division of Biological Science, Graduate School of Biology-Oriented Science and Technology, Kinki University Institute of Advanced technology, Kinki University

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Abstract

We isolated a mouse cDNA, zag1 (zygotic gene activation-associated gene 1), that has an open reading frame of 1,728-bp encoding a protein of 66.2 kDa including both a bipartite nuclear targeting sequence and a P-loop motif containing nucleoside triphosphate hydrolase motifs. Northern blot analysis of mouse tissues showed that zag1 was widely expressed but was especially prominent in the ovary and testis. RT-PCR analysis of in vitro fertilized embryos showed that the abundance of zag1 transcripts in oocytes decreased after fertilization, and zag1 mRNA was detected at 15 h post insemination (hpi) in fertilized embryos indicating that the gene was expressed at the start of zygotic gene activation at the mouse 1-cell stage. The nuclear-localization of ZAG1 protein in mouse preimplantation embryos at 15 hpi was confirmed by both subcellular analysis of enhanced green fluorescent protein (EGFP)-tagged ZAG1 and immunocytochemical analysis with anti-ZAG1 antibody. Subsequently, using yeast two-hybrid screening, we identified U2 small nuclear ribonucleoprotein B (U2B"), which is associated with pre-mRNA splicing, as a putative interacting partner of ZAG1 protein. Furthermore, knockdown of zag1 expression by an antisense DNA plasmid induced arrest and/or delay of embryonic development in injected 1-cell embryos. These results suggest that ZAG1 may be closely associated with zygotic gene expression in mouse preimplantation embryos.<br>

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