Distinct enzymatic and cellular characteristics of two secretory phospholipases A2 in the filamentous fungus Aspergillus oryzae.
Microbial secretory phospholipases A(2) (sPLA(2)s) are among the last discovered and least known members of this functionally diverse family of enzymes. We analyzed here two sPLA(2)s, named sPlaA and sPlaB, of the filamentous ascomycete Aspergillus oryzae. sPlaA and sPlaB consist of 222 and 160 amino acids, respectively, and share the conserved Cys and catalytic His-Asp residues typical of microbial sPLA(2)s. Two sPLA(2)s differ in pH optimum, Ca(2+) requirement and expression profile. The splaA mRNA was strongly upregulated in response to carbon starvation, oxidative stress and during conidiation, while splaB was constitutively expressed at low levels and was weakly upregulated by heat shock. Experiments with sPLA(2) overexpressing strains demonstrated that two enzymes produce subtly different phospholipid composition variations and also differ in their subcellular localization: sPlaA is most abundant in hyphal tips and secreted to the medium, whereas sPlaB predominantly localizes to the ER-like intracellular compartment. Both sPLA(2) overexpressing strains were defective in conidiation, which was more pronounced for sPlaB overexpressors. Although no major morphological abnormality was detected in either DeltasplaA or DeltasplaB mutants, hyphal growth of DeltasplaB, but not that of DeltasplaA, displayed increased sensitivity to H(2)O(2) treatment. These data indicate that two A. oryzae sPLA(2) enzymes display distinct, presumably non-redundant, physiological functions.
Nakahama
Tomoyuki
T
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Nakanishi
Yoshito
Y
Viscomi
Arturo R
AR
Takaya
Kohei
K
Kitamoto
Katsuhiko
K
Ottonello
Simone
S
Arioka
Manabu
M
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D003067:Coenzymes / Q000494:pharmacology; D017124:Conserved Sequence; D004721:Endoplasmic Reticulum / Q000201:enzymology; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D006863:Hydrogen-Ion Concentration; D025301:Hyphae / Q000201:enzymology; D008969:Molecular Sequence Data; D054497:Phospholipases A2, Secretory / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D003067:Coenzymes; D005656:Fungal Proteins; D010743:Phospholipids; D054497:Phospholipases A2, Secretory; D002118:Calcium