Live cell imaging of the assembly, disassembly, and actin cable-dependent movement of endosomes and actin patches in the budding yeast, Saccharomyces cerevisiae

doi:10.1083/jcb.200404173

Published 8 November 2004. doi:10.1083/jcb.200404173
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 167, Number 3, 519-530

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Article

Live cell imaging of the assembly, disassembly, and actin cable–dependent movement of endosomes and actin patches in the budding yeast, Saccharomyces cerevisiae

Thomas M. Huckaba, Anna Card Gay, Luiz Fernando Pantalena, Hyeong-Cheol Yang, and Liza A. Pon

Department of Anatomy and Cell Biology, Columbia University College of Physicians and Surgeons, New York, NY 10032

Correspondence to Liza A. Pon: lap5{at}columbia.edu

Using FM4-64 to label endosomes and Abp1p-GFP or Sac6p-GFP tolabel actin patches, we find that (1) endosomes colocalize withactin patches as they assemble at the bud cortex; (2) endosomescolocalize with actin patches as they undergo linear, retrogrademovement from buds toward mother cells; and (3) actin patchesinteract with and disassemble at FM4-64–labeled internalcompartments. We also show that retrograde flow of actin cablesmediates retrograde actin patch movement. An Arp2/3 complexmutation decreases the frequency of cortical, nonlinear actinpatch movements, but has no effect on the velocity of linear,retrograde actin patch movement. Rather, linear actin patchmovement occurs at the same velocity and direction as the movementof actin cables. Moreover, actin patches require actin cablesfor retrograde movements and colocalize with actin cables asthey undergo retrograde movement. Our studies support a mechanismwhereby actin cables serve as "conveyor belts" for retrogrademovement and delivery of actin patches/endosomes to FM4-64–labeledinternal compartments.

H.-C. Yang's present address is Department of Dental Biomaterials,Seoul National University, Seoul 110-749, Korea.

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