PpATG9 encodes a novel membrane protein that traffics to vacuolar membranes, which sequester peroxisomes during pexophagy in Pichia pastoris.
When Pichia pastoris adapts from methanol to glucose growth, peroxisomes are rapidly sequestered and degraded within the vacuole by micropexophagy. During micropexophagy, sequestering membranes arise from the vacuole to engulf the peroxisomes. Fusion of the sequestering membranes and incorporation of the peroxisomes into the vacuole is mediated by the micropexophagy-specific membrane apparatus (MIPA). In this study, we show the P. pastoris ortholog of Atg9, a novel membrane protein is essential for the formation of the sequestering membranes and assembly of MIPA. During methanol growth, GFP-PpAtg9 localizes to multiple structures situated near the plasma membrane referred as the peripheral compartment (Atg9-PC). On glucose-induced micropexophagy, PpAtg9 traffics from the Atg9-PC to unique perivacuolar structures (PVS) that contain PpAtg11, but lack PpAtg2 and PpAtg8. Afterward, PpAtg9 distributes to the vacuole surface and sequestering membranes. Movement of the PpAtg9 from the Atg9-PC to the PVS requires PpAtg11 and PpVps15. PpAtg2 and PpAtg7 are essential for PpAtg9 trafficking from the PVS to the vacuole and sequestering membranes, whereas trafficking of PpAtg9 proceeds independent of PpAtg1, PpAtg18, and PpVac8. In summary, our data suggest that PpAtg9 transits from the Atg9-PC to the PVS and then to the sequestering membranes that engulf the peroxisomes for degradation.
Chang
Tina
T
Department of Anatomy and Cell Biology, University of Florida College of Medicine, Gainesville, FL 32610-0235, USA.
Schroder
Laura A
LA
Thomson
J Michael
JM
Klocman
Amy S
AS
Tomasini
Amber J
AJ
Strømhaug
Per E
PE
Dunn
William A
WA
D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D007425:Intracellular Membranes / Q000502:physiology* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000378:metabolism*; D000432:Methanol / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D020675:Peroxisomes / Q000502:physiology* / Q000648:ultrastructure; D010843:Pichia / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014617:Vacuoles / Q000502:physiology* / Q000648:ultrastructure
D005656:Fungal Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D005947:Glucose; D000432:Methanol