A [32]P labeled DNA probe consisting of an E box was mixed with ato and/or da. Cooperative binding to the DNA was observed by EMSA.

da and ato bind cooperatively to DNA. For the DNA probe tested, neither da nor ato could bind alone.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

da and ato were mixed with [32]P labeled DNA and DNA-protein complexes observed by EMSA were used to deduce protein-protein interactions. Neither binds DNA on its own in this assay.

Two-hybrid system: yeast LexA-BD/GAL4-AD

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

The ato-da interaction is not affected by mutation of ato\'s protein kinase A phosphorylation site (S292A and S292D tested): wild-type, phospho-mimetic and phospho-null ato heterodimerize with da.