TABLE 2.
Effects of mutations in Gag or in the Ψ on interaction in the three-hybrid system
| RNA hybrida | Fusion proteina | Activationb |
|---|---|---|
| HIVΨ-MS2 | Gal4AD-HIV Gag | +++ |
| HIVΨ-MS2 | Gal4AD-HIV MA-CA | − |
| HIVΨ-MS2 | Gal4AD-HIV Gag F16A | + |
| HIVΨΔSL1,3,4-MS2 | Gal4AD-HIV Gag | ++ |
| HIVΨΔSL3,4-MS2 | Gal4AD-HIV Gag | +++ |
| HIVΨΔSL1-MS2 | Gal4AD-HIV Gag | +++ |
As in Table 1. HIVΨΔSLx, HIV-1 encapsidation signal lacking SLx, where x denotes the stem-loop number(s) as depicted in Fig. 1C. The sequence of each junction created by each deletion is as follows: ΔSL1 (deletion of nt 697 to 730 [14]), AGGAGGGG; ΔSL3 (deletion of nt 766 to 779), TTGAAAGG; ΔSL4 (deletion of nt 793 to 806), GATGAGTA. The bold letters denote the bases flanking the newly generated junctions.
As in Table 1. +++, color seen in about 15 min, turning to strong blue in about 1 h. Quantitative β-galactosidase assay yielded 420 ± 100 U for the HIVΨ-MS2/Gal4AD-HIV Gag pair. ++, color seen in about 15 min, turning blue in about 1 h. +, color seen in about 3 h, remaining fainter than the wild-type Gag after an additional 4 h of incubation. Quantitative β-galactosidase assay yielded 275 ± 100 U for both the HIVΨ-MS2/Gal4AD-HIV Gag F16A and HIVΨΔSL1,3,4-MS2/Gal4AD-HIV Gag pairs. −, no detectable color observed after overnight incubation.