TY - JOUR
T1 - Rack1 binds HIV-1 Nef and can act as a Nef-protein kinase C adaptor
AU - Gallina, A.
AU - Rossi, F.
AU - Milanesi, G.
N1 - Funding Information:
The authors thank D. Mochly-Rosen and C. Chen for the gift of plasmid pDM89, including a complete rat Rack1 gene sequence. This work was supported in part by the Italian National Institute of Health (Istituto Superiore di Sanità) III AIDS program (Grant 40B.66) and by the CNR Biotechnology Target Project. F.R. is the recipient of an Istituto Superiore di Sanità fellowship.
PY - 2001/4/25
Y1 - 2001/4/25
N2 - Nef proteins of primate immunodeficiency viruses exert pleiotropic effects, such as enhanced endocytosis of CD4 and MHC-I cell surface molecules, perturbation of signal transduction cascades, and virion infectivity enhancement. Nef function intersects that of a number of cell kinases, including C kinases (PKCs) and Src-family kinases. Here the interaction of HIV-1 Nef with Rack1 (receptor for activated C kinase 1) is reported. Nef binds the Rack1 C-terminal moiety in a yeast two-hybrid system and in cell-free pull-down assays and copurifies with in vitro translated Rack1. Nef and Rack1 partially colocalize on the trans-Golgi network and plasma membranes. The presence of Rack1 doubles Nef phosphorylation by PKCs in vitro. Our data agree with the idea that Rack1 acts as a Nef intracellular docking site, bringing Nef and PKCs together. Other signal transduction or endocytosis proteins, in particular Src-like kinases, might meet Nef by intermediation of the Rack1 adaptor.
AB - Nef proteins of primate immunodeficiency viruses exert pleiotropic effects, such as enhanced endocytosis of CD4 and MHC-I cell surface molecules, perturbation of signal transduction cascades, and virion infectivity enhancement. Nef function intersects that of a number of cell kinases, including C kinases (PKCs) and Src-family kinases. Here the interaction of HIV-1 Nef with Rack1 (receptor for activated C kinase 1) is reported. Nef binds the Rack1 C-terminal moiety in a yeast two-hybrid system and in cell-free pull-down assays and copurifies with in vitro translated Rack1. Nef and Rack1 partially colocalize on the trans-Golgi network and plasma membranes. The presence of Rack1 doubles Nef phosphorylation by PKCs in vitro. Our data agree with the idea that Rack1 acts as a Nef intracellular docking site, bringing Nef and PKCs together. Other signal transduction or endocytosis proteins, in particular Src-like kinases, might meet Nef by intermediation of the Rack1 adaptor.
UR - http://www.scopus.com/inward/record.url?scp=0035946309&partnerID=8YFLogxK
U2 - 10.1006/viro.2001.0855
DO - 10.1006/viro.2001.0855
M3 - Article
SN - 0042-6822
VL - 283
SP - 7
EP - 18
JO - Virology
JF - Virology
IS - 1
ER -