Microautophagy is promoted after nutrient starvation and inactivation of target of rapamycin complex 1 (TORC1) kinase. Invagination of vacuolar membranes by endosomal sorting complex required for transport (ESCRT) is required for microautophagy. Vps27, a subunit of ESCRT-0, is recruited onto vacuolar membranes via dephosphorylation after TORC1 inactivation. Here, we showed that Hsel, another ESCRT-0 subunit, is also recruited onto vacuolar membranes after TORC1 inactivation, promoting formation of ESCRT-0 complex on vacuolar membranes. Hsel recruitment was dependent on Vps27, whereas Vps27 recruitment was independent of Hsel. Not only Vps27 but also Hsel was required for ESCRT-III recruitment onto vacuolar membranes and microautophagy induction after TORC1 inactivation. This study revealed that ESCRT-0 (Vps27-Hsel) complex formation on vacuolar membranes is important for microautophagy inactivation after TORC1 inactivation. (C) 2019 Elsevier Inc. All rights reserved.