C-box mutation results in increased RNA encapsidation. (A) A schematic of the radioactive probe and predicted protected fragments based on the presence of a wild-type or mutant G-box. (B) RNase protection assay of intracellular RNAs obtained from duplicate experiments. DNA templates used for each transfection are listed below each lane (lanes 1–8). Lanes 9 and 10 contain yeast RNA used as a nonspecific target control, incubated with or without RNases, respectively. (C) RNase protection assay of virion RNAs produced from duplicate HIV-2 DNA transfected COS-7 cell experiments. DNA templates used for each transfection are listed below each lane (lanes 1–8). (D) Quantification of B and C. RNA levels were normalized to wild-type levels, set at 100% for both intracellular and virion samples. Identities of RNA source, intracellular or virion, and DNA templates used for each transfection are listed on the x-axis. The y-axis represents gag and gag-pol RNA levels in percent. The y-axis error bars indicate standard error for the two experiments.
