Analysis of interaction of 16.4.1 with Rev. (A) Schematic overview of domains of Rev. Locations of functional regions are taken from . Numbering of amino acids (aa) is based on the Rev sequence of HIV-1 isolate HXB-2. MI and MII: Regions that direct multimerization of Rev. NLS: nuclear localization signal; NES: nuclear export signal. (B) Identification of amino acid positions in Rev required for interaction with 16.4.1. Bait proteins containing Rev or various mutants of Rev were analysed for interaction with 16.4.1-prey. Numbers indicate positions of amino acid changes in Rev mutants. Mutations are as follows: RevM4: Y23 to D23, S25 to D25, N26 to L26 ; Rev M10BL: L78 to D78, E79 to L79 and insertion of EDLP between L81 and T82 ; Rev M5: R38 to D38, R39 to L39 ; RevSLT40: I59 to D59, L60 to D59 . Interaction is indicated by growth of yeast transformants under selective conditions (≥ 500 transformants per plate). Results represent four independent experiments. The red box marks the amino acid positions in Rev required for interaction with 16.4.1 and the location of the putative 16.4.1-interaction region of Rev (aa 38 to 60). (C) Identification of regions of 16.4.1 required for interaction with Rev. Prey proteins containing various regions of the 16.4.1 domain were analysed for interaction with Rev-bait. Interaction was analysed by growth of yeast transformants under selective conditions. Results represent three independent experiments. +++ 700–1400 transformants per plate; + 200–400 transformants per plate; – no transformants. 16.4.1 regions comprising amino acids 2 to 133 and 39 to 171 interacted with Rev with similar efficiency as full-length 16.4.1, whereas regions 2 to 73 and 74 to 171 showed weaker interaction. No interaction was observed with regions 2 to 38 and 134 to 171. The red arrow indicates the putative Rev-interaction region of 16.4.1 (aa 39 to 133).
Kramer-Hämmerle et al. BMC Cell Biology 2005 6:20 doi:10.1186/1471-2121-6-20