Oryzapsins, the orthologs of yeast yapsin in Aspergillus oryzae, affect ergosterol synthesis

Natsuno Shimizu, Tamaki Katagiri, Akira Matsumoto, Yoshihiko Matsuda, Hiroshi Arai, Nobumitsu Sasaki, Keietsu Abe, Toru Katase, Hiroki Ishida, Ken Ichi Kusumoto, Michio Takeuchi, Youhei Yamagata

    Research output: Contribution to journalArticlepeer-review

    Abstract

    The oryzapsin genes opsA and opsB in Aspergillus oryzae encoding glycosylphosphatidylinositol (GPI)-anchored aspartic endopeptidase are homologs of Saccharomyces cerevisiae yapsins. We recently found another homolog, opsC, in the A. oryzae genome database, which was suggested to be a pseudogene. However, the profiles and roles of the proteins encoded by these genes have not yet been clarified. Toward this end, we first produced opsA- and opsB-overexpression strains and performed enzymatic analyses, revealing that OpsA and OpsB can attack sites other than the carboxyl-terminal peptide bonds of basic amino acids. Moreover, OpsA and OpsB were confirmed to bind to the cell membrane with a GPI anchor. Second, opsA and opsB single-deletion and double-deletion strains (ΔopsA, ΔopsB, and ΔopsAΔopsB) were constructed to explore the expected roles of oryzapsins in cell wall synthesis, similar to the role of yapsins. The transcription level of mpkA in the cell wall integrity pathway was increased in ΔopsB and ΔopsAΔopsB strains, suggesting that OpsB might be involved in processing cell wall synthesis-related proteins. Treatment with an ergosterol biosynthesis inhibitor reduced the growth of the ΔopsAΔopsB strain. Moreover, the mRNA levels of Aoerg1, Aoerg3-1, Aoerg3-2, Aoerg7b, Aoerg11, and Aohmg1,2 showed a decreasing tendency in the ΔopsAΔopsB strain, and the ergosterol content in the membrane was reduced in the ΔopsAΔopsB strain. These results suggest that oryzapsins exist in the cell membrane and play roles in the formation of cell membranes. This is the first report of the involvement of GPI-anchored aspartic endopeptidases in ergosterol biosynthesis. Key points • The oryzapsins have wider substrate specificity than yaspins in S. cerevisiae. • Unlike the yapsins, the oryzapsins might not be involved in the main structure synthesis of the cell wall. • The oryzapsins would be involved in ergosterol biosynthesis.

    Original languageEnglish
    Pages (from-to)8481-8494
    Number of pages14
    JournalApplied Microbiology and Biotechnology
    Volume105
    Issue number21-22
    DOIs
    Publication statusPublished - 2021 Nov

    Keywords

    • Aspartic endopeptidase
    • Aspergillus oryzae
    • Cell membrane
    • Cell wall integrity pathway
    • Ergosterol synthesis
    • GPI-anchored protein

    ASJC Scopus subject areas

    • Biotechnology
    • Applied Microbiology and Biotechnology

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