TY - JOUR
T1 - Oryzapsins, the orthologs of yeast yapsin in Aspergillus oryzae, affect ergosterol synthesis
AU - Shimizu, Natsuno
AU - Katagiri, Tamaki
AU - Matsumoto, Akira
AU - Matsuda, Yoshihiko
AU - Arai, Hiroshi
AU - Sasaki, Nobumitsu
AU - Abe, Keietsu
AU - Katase, Toru
AU - Ishida, Hiroki
AU - Kusumoto, Ken Ichi
AU - Takeuchi, Michio
AU - Yamagata, Youhei
N1 - Funding Information:
This work was partially supported by the Program for the Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN). The authors thank Dr. Junichiro Marui (Applied Microbiology Division, National Food Research Organization) for kindly providing A. oryzae ΔpyrG strain.
Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
PY - 2021/11
Y1 - 2021/11
N2 - 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.
AB - 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.
KW - Aspartic endopeptidase
KW - Aspergillus oryzae
KW - Cell membrane
KW - Cell wall integrity pathway
KW - Ergosterol synthesis
KW - GPI-anchored protein
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U2 - 10.1007/s00253-021-11639-7
DO - 10.1007/s00253-021-11639-7
M3 - Article
C2 - 34668983
AN - SCOPUS:85117286303
VL - 105
SP - 8481
EP - 8494
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
SN - 0175-7598
IS - 21-22
ER -