title: Subcellular localization of acyl-CoA binding protein in Aspergillus oryzae is regulated by autophagy machinery.
abstract: In eukaryotic cells, acyl-CoA binding protein (ACBP) is important for cellular activities, such as in lipid metabolism. In the industrially important fungus Aspergillus oryzae, the ACBP, known as AoACBP, has been biochemically characterized, but its physiological function is not known. In the present study, although we could not find any phenotype of AoACBP disruptants in the normal growth conditions, we examined the subcellular localization of AoACBP to understand its physiological function. Using an enhanced green fluorescent protein (EGFP)-tagged AoACBP construct we showed that AoACBP localized to punctate structures in the cytoplasm, some of which moved inside the cells in a microtubule-dependent manner. Further microscopic analyses showed that AoACBP-EGFP co-localized with the autophagy marker protein AoAtg8 tagged with red fluorescent protein (mDsRed). Expression of AoACBP-EGFP in disruptants of autophagy-related genes revealed aggregation of AoACBP-EGFP fluorescence in the cytoplasm of Aoatg1, Aoatg4 and Aoatg8 disruptant cells. However, in cells harboring disruption of Aoatg15, which encodes a lipase for autophagic body, puncta of AoACBP-EGFP fluorescence accumulated in vacuoles, indicating that AoACBP is transported to vacuoles via the autophagy machinery. Collectively, these results suggest the existence of a regulatory mechanism between AoACBP localization and autophagy.
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Kouhei
authors.initials: K
authors.lastname: Kawaguchi
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kikuma
authors.affiliation: Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan. Electronic address: y.higuchi@agr.kyushu-u.ac.jp.
authors.forename: Yujiro
authors.initials: Y
authors.lastname: Higuchi
authors.affiliation: Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Takegawa
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: k-kitamoto@nichiyaku.ac.jp.
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Kitamoto
mesh_terms: D001236:Aspergillus oryzae / Q000166:cytology / Q000378:metabolism*; D001343:Autophagy / Q000502:physiology*; D000071190:Autophagy-Related Protein 8 Family / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D026261:Diazepam Binding Inhibitor / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D000071190:Autophagy-Related Protein 8 Family; D026261:Diazepam Binding Inhibitor; D005656:Fungal Proteins; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein
keywords: Acyl-CoA binding protein; Aspergillus oryzae; Autophagy; Microtubule