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Published online 24 January 2005. doi:10.1083/jcb.200407113
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 168, Number 3, 477-488
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Article

 Clathrin- and caveolin-1–independent endocytosis : entry of simian virus 40 into cells devoid of caveolae



Eva-Maria Damm1, Lucas Pelkmans2, Jürgen Kartenbeck3, Anna Mezzacasa1, Teymuras Kurzchalia2, and Ari Helenius1

1 Institute of Biochemistry, Swiss Federal Institute of Technology Zürich (ETHZ), CH-8093 Zürich, Switzerland
2 Max Planck Institute for Molecular Cell Biology and Genetics, D-01307 Dresden, Germany
3 German Cancer Research Center (DKFZ) Heidelberg, D-69120 Heidelberg, Germany

Correspondence to Ari Helenius: ari.helenius{at}bc.biol.ethz.ch

Simian Virus 40 (SV40) has been shown to enter host cells by caveolar endocytosis followed by transport via caveosomes to the endoplasmic reticulum (ER). Using a caveolin-1 (cav-1)–deficient cell line (human hepatoma 7) and embryonic fibroblasts from a cav-1 knockout mouse, we found that in the absence of caveolae, but also in wild-type embryonic fibroblasts, the virus exploits an alternative, cav-1–independent pathway. Internalization was rapid (t1/2 = 20 min) and cholesterol and tyrosine kinase dependent but independent of clathrin, dynamin II, and ARF6. The viruses were internalized in small, tight-fitting vesicles and transported to membrane-bounded, pH-neutral organelles similar to caveosomes but devoid of cav-1 and -2. The viruses were next transferred by microtubule-dependent vesicular transport to the ER, a step that was required for infectivity. Our results revealed the existence of a virus-activated endocytic pathway from the plasma membrane to the ER that involves neither clathrin nor caveolae and that can be activated also in the presence of cav-1.

Abbreviations used in this paper: AF, Alexafluor; BFA, Brefeldin A; cav-1, caveolin-1; cav-1WT, cav-1 wild-type; HuH7, human hepatoma 7; KO, knockout; LY, Lucifer yellow; MOI, multiplicity of infection; SFV, Semliki Forest virus; SV40, simian virus 40; Tf, transferrin.


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