UDP-N-acetylglucosamine-dolichyl-phosphate N-acetylglucosaminephosphotransferase is indispensable for oogenesis, oocyte-to-embryo transition, and larval development of the nematode Caenorhabditis elegans

Nanako Kanaki, Ayako Matsuda, Katsufumi Dejima, Daisuke Murata, Kazuko H. Nomura, Takashi Ohkura, Keiko Gengyo-Ando, Sawako Yoshina, Shohei Mitani, Kazuya Nomura

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

N-linked glycosylation of proteins is the most common post-translational modification of proteins. The enzyme UDP-N-acetylglucosamine-dolichyl-phosphate N-acetylglucosaminephosphotransferase (DPAGT1) catalyses the first step of N-glycosylation, and DPAGT1 knockout is embryonic lethal in mice. In this study, we identified the sole orthologue (algn-7) of the human DPAGT1 in the nematode C. Elegans. The gene activity was disrupted by RNAi and deletion mutagenesis, which resulted in larval lethality, defects in oogenesis and oocyte-to-embryo transition. Endomitotic oocytes, abnormal fusion of pronuclei, abnormal AB cell rotation, disruption of permeation barriers of eggs, and abnormal expression of chitin and chitin synthase in oocytes and eggs were the typical phenotypes observed. The results indicate that N-glycosylation is indispensable for these processes. We further screened an N-glycosylated protein database of C. Elegans, and identified 456 germline-expressed genes coding N-glycosylated proteins. By examining RNAi phenotypes, we identified five germline-expressed genes showing similar phenotypes to the algn- 7 (RNAi) animals. They were ribo-1, stt-3, ptc-1, ptc-2, and vha-19. We identified known congenital disorders of glycosylation (CDG) genes (ribo-1 and stt-3) and a recently found CDG gene (vha-19). The results show that phenotype analyses using the nematode could be a powerful tool to detect new CDG candidate genes and their associated gene networks.

Original languageEnglish
Pages (from-to)163-178
Number of pages16
JournalGlycobiology
Volume29
Issue number2
DOIs
Publication statusPublished - 2018 Nov 16
Externally publishedYes

Keywords

  • ALG7
  • DPAGT1
  • N-glycosylated proteins
  • RNAi
  • gene knockout

ASJC Scopus subject areas

  • Biochemistry

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