Distinct tra trafficking pathways mediate Nef-induced and clathrin-dependent major histocompatibility complex class I down-regulation

The human immunodeficiency virus type 1 Nef protein alters the post-Golgi s tages of major histocompatibility complex class I (MHC-I) biogenesis. Presu med mechanisms involve the disclosure of a cryptic tyrosine based sorting s ignal (YSQA) located in the cytoplasmic tail of HLA-A and -B heavy chains. We changed this signal for a prototypic sorting motif (YSQI or YSQL). Modif ied HLA-A2 molecules, termed A2-endo, displayed constitutively low surface levels and accumulated in a region close to or within the Golgi apparatus, a behavior reminiscent of wild-type HLA-A2 in Nef-expressing cells. However , several lines of evidence indicate that the action of prototypic signals on MHC-I trafficking differs from that of Nef. Internalization of surface A 2-endo was more rapid and was associated with efficient recycling to the su rface. A transdominant-negative mutant of dynamin-1 inhibited A2-endo const itutive internalization and Nef-induced CD4 down-regulation, whereas it did not affect the activity of Nef on MHC-I. Moreover, trafficking of A2 endo was still affected by the viral protein, indicating additive effects of pro totypic signals and Nef, Therefore, distinct trafficking pathways regulate clathrin-dependent and Nef-induced MHC-I modulation.