A Myosin I Is Involved in Membrane Recycling from Early Endosomes

doi:10.1083/jcb.150.5.1013

Published online 5 September 2000. doi:10.1083/jcb.150.5.1013

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© The Rockefeller University Press, 0021-9525/2000/9/1013/ $5.00
The Journal of Cell Biology, Volume 150, Number 5, September 4, 2000 1013-1026

Original Article

A Myosin I Is Involved in Membrane Recycling from Early Endosomes

Eva M. Neuhaus^a and Thierry Soldati^a
^a Department of Molecular Cell Research, Max-Planck-Institute for Medical Research, D-69120 Heidelberg, Germany

Correspondence to: Thierry Soldati, Department of Molecular Cell Research, Max-Planck-Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany. Tel:49-6221-486407 Fax:49-6221-486325 E-mail:soldati{at}mpimf-heidelberg.mpg.de.

Geometry-based mechanisms have been proposed to account forthe sorting of membranes and fluid phase in the endocytic pathway,yet little is known about the involvement of the actin–myosincytoskeleton. Here, we demonstrate that Dictyostelium discoideummyosin IB functions in the recycling of plasma membrane componentsfrom endosomes back to the cell surface. Cells lacking MyoB(myoA^-/B^-, and myoB^- cells) and wild-type cells treated withthe myosin inhibitor butanedione monoxime accumulated a plasmamembrane marker and biotinylated surface proteins on intracellularendocytic vacuoles. An assay based on reversible biotinylationof plasma membrane proteins demonstrated that recycling of membranecomponents is severely impaired in myoA/B null cells. In addition,MyoB was specifically found on magnetically purified early pinosomes.Using a rapid-freezing cryoelectron microscopy method, we observedan increased number of small vesicles tethered to relativelyearly endocytic vacuoles in myoA^-/B^- cells, but not to laterendosomes and lysosomes. This accumulation of vesicles suggeststhat the defects in membrane recycling result from a disorderedmorphology of the sorting compartment.

Key Words: Dictyostelium, endocytosis, membrane recycling, myosins, sorting

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