Fig. 7
From: Myristoylation drives dimerization of matrix protein from mouse mammary tumor virus
Mutations L11N, F12N and V15N in MA domain (MA-3N) reduce extracellular MMTV particle production. a Pulse-chase analysis of immature MMTV particles produced in transfected 293T cells. Cells expressed wild-type MMTV (WT) and the virus with the mutation in MA domain of Gag (MA-3N); M mock-transfected cells. Cells were labeled for 1 h with [35S]methionine and [35S]cysteine (lanes 1–3) and then chased for 12 h (lanes 4–5). At both time points, cells were harvested, lysed, and subjected to immunoprecipitation. Released particles (lanes 6–7) were collected from the culture media by ultracentrifugation through a 20 % sucrose cushion. Viral proteins were immunoprecipitated with polyclonal rabbit anti-CA serum, separated on a 10 % SDS-PAGE gel, and subjected to phosphorimager analysis. b Comparative analysis of released particle production. The efficiency was expressed as a share of the signal from the released particulate Gag from the total amount of Gag synthesized. Mutant MA-3N is shown relative to the WT (set to 1). Each bar represents the average of results from three independent experiments, and standard error of the mean is shown