Systematic Interaction Mapping between 46 Circadian Clock Proteins and Associated Components.

<p>(A) Matrix based high-throughput yeast-two-hybrid interaction screen. (B) CLOCK interactors: Mating controls (top left); upon PPI reporter genes are activated (top middle: HIS, URA for growth selection, top right: lacZ for β-galactosidase activity). Bottom: Detected interactions with CLOCK; red lines: interactions previously discovered in yeast (see also <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003398#pgen.1003398.s001" target="_blank">Figure S1</a>). (C) Clock protein interaction matrix. Circles: interactions between two components not differentiating between bait and prey configuration. (D) Validation of new CLOCK and BMAL1 interactions in mammalian cells. HEK293 cells expressing CLOCK- or BMAL1-luciferase fusions were transfected with MYC-tagged components. Luciferase activity in anti-MYC co-immunoprecipitates is presented for one representative result of at least two independent experiments with similar results (for method and input controls also see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003398#pgen.1003398.s002" target="_blank">Figure S2</a>). MYC-β-galcactosidase fusions served as negative, MYC-BMAL1 and MYC-CRY1 as positive controls, respectively.