Visualization of the endocytic pathway in the filamentous fungus Aspergillus oryzae using an EGFP-fused plasma membrane protein.
Endocytosis is an important process for cellular activities. However, in filamentous fungi, the existence of endocytosis has been so far elusive. In this study, we used AoUapC-EGFP, the fusion protein of a putative uric acid-xanthine permease with enhanced green fluorescent protein (EGFP) in Aspergillus oryzae, to examine whether the endocytic process occurs or not. Upon the addition of ammonium into the medium the fusion protein was internalized from the plasma membrane. The internalization of AoUapC-EGFP was completely blocked by sodium azide, cold, and cytochalasin A treatments, suggesting that the internalization possesses the general features of endocytosis. These results demonstrate the occurrence of endocytosis in filamentous fungi. Moreover, we discovered that the endosomal compartments appeared upon the induction of endocytosis and moved in a microtubule-dependent manner.
Higuchi
Yujiro
Y
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Nakahama
Tomoyuki
T
Shoji
Jun-Ya
JY
Arioka
Manabu
M
Kitamoto
Katsuhiko
K
D000199:Actins / Q000737:chemistry; D001236:Aspergillus oryzae / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D003080:Cold Temperature; D003572:Cytochalasins / Q000494:pharmacology; D004705:Endocytosis; D011992:Endosomes; D005656:Fungal Proteins / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000737:chemistry; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000737:chemistry / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D019810:Sodium Azide / Q000494:pharmacology; D013997:Time Factors
D000199:Actins; D003572:Cytochalasins; D005656:Fungal Proteins; D026901:Membrane Transport Proteins; D000644:Quaternary Ammonium Compounds; D011993:Recombinant Fusion Proteins; C083852:UAPA protein, Aspergillus nidulans; C485184:enhanced green fluorescent protein; C031218:cytochalasin A; D049452:Green Fluorescent Proteins; D019810:Sodium Azide