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. Author manuscript; available in PMC: 2009 Apr 25.
Published in final edited form as: Mol Cell. 2008 Apr 25;30(2):227–238. doi: 10.1016/j.molcel.2008.03.015

Figure 3.

Figure 3

GGA overexpression inhibits HIV-1 Gag binding to membrane and an HIV-1 proviral clone bearing a foreign membrane-targeting signal is resistant to the inhibitory effects of GGA overexpression. (A) HeLa cells were transfected with pNL4-3/PR− along with control vector or GGA or Tsg101 (TSG-F) expression constructs. Cells were pulse-labeled with [35S]Met/Cys for 5 min and chased for 15 min in cold medium. Membrane (M) and non-membrane (NM) fractions were separated by membrane flotation centrifugation. The individual fractions were immunoprecipitated with HIV-Ig after denaturation. (B) Quantitation of data; ± SD, n=3. Position of Pr55Gag is indicated (Pr55). (C) Schematic depiction of the Fyn10deltaMA construct, with myristic (Myr) and palmitic (Palm) acid moieties shown. (D) Effect of GGA overexpression on the release of WT or Fyn10deltaMA virus. HeLa cells were transfected with either WT pNL4-3 or Fyn10deltaMA proviral construct along with GGA or Tsg101 (TSG-F) expression plasmids. Cell and virus lysates were radioimmunoprecipitated with HIV-Ig. Means ± SD, n=3.