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© The Rockefeller University Press, 0021-9525/1997//1007 $5.00
The Journal of Cell Biology, Volume 136, Number 5, , 1997 1007-1021

Article

Microtubule-mediated Transport of Incoming Herpes Simplex Virus 1 Capsids to the Nucleus



Beate Sodeik, Melanie W. Ebersold, and Ari Helenius

Yale University School of Medicine, Department of Cell Biology, New Haven, Connecticut 06520-8002

Herpes simplex virus 1 fuses with the plasma membrane of a host cell, and the incoming capsids are efficiently and rapidly transported across the cytosol to the nuclear pore complexes, where the viral DNA genomes are released into the nucleoplasm. Using biochemical assays, immunofluorescence, and immunoelectron microscopy in the presence and absence of microtubule depolymerizing agents, it was shown that the cytosolic capsid transport in Vero cells was mediated by microtubules. Antibody labeling revealed the attachment of dynein, a minus end–directed, microtubule-dependent motor, to the viral capsids. We propose that the incoming capsids bind to microtubules and use dynein to propel them from the cell periphery to the nucleus.

Abbreviations used in this paper: CH, cycloheximide; GA, glutaraldehyde; HSV-1, herpes simplex virus 1; MOI, multiplicity of infection; MT, microtubule; MTOC, microtubule organizing center; PFA, paraformaldehyde; PFU, plaque-forming unit; PI, postinfection; TX-100, Triton X-100.

B. Sodeik thanks Drs. Gary Cohen and Roselyn Eisenberg for their generous hospitality in their labs and many helpful suggestions. Victor Stolc is acknowledged for his contribution to the immunofluorescence studies. We also thank Lisa Hartnell, Linda Iadarola, Philippe Male, and Paul Webster of the Center of Cell Imaging in the Cell Biology Department at Yale University for teaching many aspects of EM, and Henry Tan for his superb photographic work. We are grateful to Drs. Gary Whittaker, Jani Simons, Päivi Ojala, Philippe Pierre, and Urs Greber for many stimulating discussions and critical reading of the manuscript.

The work was supported by a National Institutes of Health grant to A. Helenius (AI18599) and a long-term postdoctoral EMBO fellowship to B. Sodeik (ALTF 254-1993).

Address all correspondence to Beate Sodeik, Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520-8002. Tel.: (203) 737-2612. Fax: (203) 785-7226. e-mail: sodeik{at}biomed.med.yale.edu


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