Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

The cry-tim interaction was observed in cells kept in constant darkness and in cells exposed to light for one hour, but not in cells exposed to continuous light.

Two-hybrid system: yeast LexA-BD/VP16-AD

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

This interaction was observed only in cells exposed to constant light (not constant darkness).

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast LexA-BD/B42-AD

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast LexA-BD/B42-AD, interaction depends on exposure to light.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast LexA-BD/B42-AD

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

The cry-tim interaction is only detected when yeast cells are grown in light.

Two-hybrid system: yeast LexA-BD/B42-AD

Positive control.

Interaction in vitro; bait produced by chemical synthesis; prey produced as a recombinant fusion protein in baculovirus-infected insect cell system.

Treatment of cry with EDTA, which reduces the bound-FAD chromophore to anionic semiquinone (ASQ), bypasses the requirement of light in the cry-tim interaction.

The cry W536F mutant was used for the binding assay as it exhibits the sharpest light-induced conformational changes, as determined by partial trypsin digestion.

Wildtype cry binds tim preferentially in the dark while a version of cry with the C-terminal tail (CTT) deleted binds tim equally well in light and dark.

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

Wild-type cry interaction with tim increased with exposure to light, but does not have an effect on cry delta-CTT, aa 377 or aa 378 mutants\' interaction with tim. However, cry delta-CTT and aa 377 mutants had an overall increased interaction with tim in both light and dark conditions.

Source was cell extract of S2 cell line; bait and prey produced from transfected construct.