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Originally published as MBC in Press, 10.1091/mbc.E03-08-0578 on November 14, 2003

Vol. 15, Issue 1, 323-331, January 2004

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HIV Nef-mediated Major Histocompatibility Complex Class I Down-Modulation Is Independent of Arf6 Activity

Jakob E. Larsen *, Ramiro H. Massol *, Thomas J. F. Nieland, and Tomas Kirchhausen {dagger}

Department of Cell Biology and The Center for Blood Research, Harvard Medical School, Boston, Massachusetts 02115

Submitted August 11, 2003; Revised September 3, 2003; Accepted September 10, 2003
Monitoring Editor: Juan Bonifacino

HIV Nef has a number of important biological effects, including the down-modulation of several immunological important molecules (CD4, major histocompatibility complex [MHC] class I). Down-modulation of CD4 seems to be via clathrin-dependent endocytosis, whereas down-modulation of MHC class I remains unexplained. Several mutant proteins, including mutations in the small GTPase Arf6, have been used to probe membrane traffic pathways. One such mutant has recently been used to propose that Nef acts through Arf6 to activate the endocytosis of MHC class I. Here, we show that MHC class I down-modulation is unaffected by other Arf6 mutants that provide more specific perturbations in the GDP-GTP cycling of Arf6. Inhibition of phosphatidylinositol-3-phosphate kinase, an upstream activator of Arf6, also had no effect on the internalization step, but its activity is required to direct MHC class I to the trans-Golgi network. We conclude that the apparent Arf6 dependency of Nef-mediated MHC class I down-modulation is due to nonspecific perturbations in membrane traffic.

Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E03–08–0578. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E03–08–0578.

* These authors contributed equally to this work.

{dagger} Corresponding author. E-mail address: kirchhausen{at}crystal.harvard.edu.




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