Figure 1—figure supplement 1. | RNA-directed remodeling of the HIV-1 protein Rev orchestrates assembly of the Rev–Rev response element complex

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RNA-directed remodeling of the HIV-1 protein Rev orchestrates assembly of the Rev–Rev response element complex

Figure 1—figure supplement 1.

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University of California, San Francisco, United States
Figure 1—figure supplement 1.
Download figureOpen in new tabFigure 1—figure supplement 1. Rev and RRE constructs.

(A) Rev dimer structure (Daugherty et al., 2010b) with the higher-order oligomerization disrupting mutations, L12S and L60R shown in red. Also shown is the surface-entropy reducing mutation, E47A in blue or green. (B) RRE IIB40 is a derivative of the stem II three-helix junction and contains two adjacent Rev-binding sites (red). (C) Gel-shift assays comparing binding of 32P-labeled RRE-stem II or IIB40 to Rev. Free, F, monomer, M and dimer, D complexes are indicated. A doublet/smeared band is observed for the monomer complex with both RNAs and is indicative of conformational heterogeneity. (D) Binding curves calculated from gel-shift assays in (C). Apparent dissociation constants, Kd and Hill coefficient (n) were determined using the equation: Fraction of RNA bound = [Rev]n/(Kdn + [Rev]n) as mean ± s.d. of two replicates.

DOI: http://dx.doi.org/10.7554/eLife.04120.004