Vacuolar processing enzyme translocates to the vacuole through the autophagy pathway to induce programmed cell death

Abstract

The caspase-like vacuolar processing enzyme (VPE) is a key factor in programmed cell death (PCD) associated with plant stress responses. Growth medium lacking a carbon source and dark conditions caused punctate labeling of 35S::VPE1-GFP (StVPE1-GFP) in potato leaves. Carbon starvation of BY-2 cells induced higher VPE activity and PCD symptoms. Growing VPE-RNAi BY-2 cells without sucrose reduced VPE activity and prevented PCD symptoms. During extended exposure to carbon starvation, VPE expression and activity levels peaked, with a gradual increase in BY-2 cell death. Histological analysis of StVPE1-GFP in BY-2 cells showed that carbon starvation induces its translocation from the endoplasmic reticulum to the central vacuole, through tonoplast engulfment. Exposure of BY-2 culture to the autophagy inhibitor concanamycin A caused autophagic bodies accumulation in the cell vacuole. Such accumulation did not occur in the presence of 3-methyladenine, an inhibitor of early-stage autophagy. BY-2 cells constitutively expressing StATG8IL-RFP, an autophagosome marker, showed colocalization with the StVPE1-GFP protein in the cytoplasm and vacuole. RNAi silencing of the core autophagy component ATG4 in BY-2 cells reduced VPE activity and cell death. These results are the first to suggest that VPE translocates to the cell vacuole through the autophagy pathway, leading to PCD.\n\nOne sentence summaryCarbon starvation induced programmed cell death by trafficking vacuolar processing enzyme through the autophagy pathway to the vacuole.