Supplemental Materials
This article contains the following supporting material:
- Supplemental Materials
- Movie01 - Movie 1. Cdc8p-decorated actin filaments support the processive movement of single Myo52p-Qdot molecules. Left: Failure of Myo52p-HMM-Qdots (green) to move processively on bare actin filaments (red) in the presence of 2 mM ATP. Right: In contrast, Myo52p-HMM-Qdots can move processively on Cdc8pdecorated actin filaments. Data were collected using TIRF microscopy with a capture rate of 30 frames/s and are played back at this speed. Scale bar: 2 μm.
- Movie02 - Movie 2. Cdc8p-decorated actin filaments support the processive movement of single Myo52p-Cy3 molecules. Cy3-labeled Myo52p-HMM (green) moving processively on actin-Cdc8 filaments (red) in the presence of 2 mM ATP. Data were collected using TIRF microscopy with a capture rate of 5 frames/s and are played back at this speed. Scale bar: 2 μm.
- Movie03 - Movie 3. Processive movement of Myo52p molecules on bare actin and Cdc8p-decorated actin filaments at low concentrations of ATP. Left: A Myo52p-HMM-Qdot (green) moving processively on bare actin filaments (red) in the presence of 10 μM ATP. Right: Myo52p-HMM-Qdots moving processively on Cdc8p-decorated actin filaments in 10 μM ATP. Data were collected using TIRF microscopy with a capture rate of 1 frame/s and are played back at 30 frames/s (sped-up 30x). Scale bar: 2 μm.
- Movie04 - Movie 4. Cdc8p-decorated actin filaments support movement of Qdots by multiple Myo52p molecules. Left: Failure of Qdots (green) bound by multiple Myo52p-HMM motors to move along bare actin filaments (red) in the presence of 2 mM ATP. Right: In contrast, Qdots bound by multiple Myo52p- HMM motors can move processively on Cdc8p-decorated actin filaments. Data were collected using TIRF microscopy with a capture rate of 30 frames/s and are played back at this speed. Scale bar: 2 μm.
- Movie05 - Movie 5. The directed motility of Myo52p molecules in the cell does not rely on the C-terminal cargobinding domain. In vivo directed motility of Myo52pΔCBD-3xGFP particles (green) made from clusters of molecules lacking their C-terminal cargo-binding domains in the myo52ΔCBD-3xGFP strain. Data were collected using epi-fluorescence microscopy with a capture rate of 10 frames/s; played back at 100 frames/s (sped up 10x).