Model for how Vac17 uses a handhold to stabilize its interaction with Myo2 and initiate vacuole movement into small buds. (A) Vacuole inheritance occurs concurrently with bud emergence or in small buds. The Myo2–Vac17–Vac8 complex localizes to a region of the vacuole near the bud site, and a portion of the vacuole is subsequently moved into the bud. (B) The proper timing of vacuole transport depends on stable interactions between Myo2 and Vac17. This is achieved by Vac17 engaging Myo2 at two distinct interfaces via its Vac1(H) and Vac17(MBD) regions. (C and D) In the vac17ΔH mutant, the initiation of vacuole movement is significantly delayed. This is likely due to a loss of stable interactions with Myo2 and highlights the critical role of the handhold mechanism in ensuring timely vacuole transport. (E) Vacuole inheritance eventually occurs in large buds of the vac17ΔH mutant. This is likely due to a defect in the turnover of Vac17, resulting in an abnormal accumulation of Vac17 on the vacuole membrane. (F) We speculate that this excess availability of Vac17 compensates for the partial defect in vac17ΔH by promoting increased recruitment of Myo2 to the vacuole or by increasing the occupancy of Vac17(MBD) for Myo2. MBD, Myo2-binding domain.