loqs coimmunoprecipitates with Dcr-1 but not Dcr-2.

Source was cell extract of S2 cell line; bait produced from endogenous gene; prey produced from endogenous gene.

Purified proteins were incubated with [32]P labeled let-7 pre-miRNA and protein:RNA complex formation was monitored by gel shift.

Dcr-1 and loqs form a higher molecular weight RNA:protein complex with let-7 than just Dcr-1 alone. loqs alone does not bind let-7 pre-miRNA in this assay.

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

Dcr-1 coimmunoprecipitates with loqs (r3d1) but not r2d2.

Source was cell extract of S2 cell line; bait produced from endogenous gene; prey produced from endogenous gene.

Source was cell extract of S2 cell line; proteins produced from endogenous genes.

Extract was fractionated by sequential ammonium sulfate precipitation and phenyl-sepharose, Mono S, Mono Q and hydroxyapatite chromatography.

Dcr-1 and loqs (L isoform) cofractionate together and with an miRNA-generating activity through several chromatographic steps.

Dcr-1 coimmunoprecipitates with both short and long isoforms of tagged transfected loqs. By contrast, endogenous long loqs isoforms preferentially coimmunoprecipitate with Dcr-1.

Dcr-1, but not Dcr-2, coimmunoprecipitates with loqs.

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

Purified proteins were mixed and subjected to sedimentation analysis.

When mixed with loqs, Dcr-1 migrates at a position consistent with the formation of a Dcr-1-loqs heterodimer.

Interaction in vitro; bait produced as a recombinant fusion protein in baculorvirus-infected Sf21 cell system; prey produced as a recombinant fusion protein in baculorvirus-infected Sf21 cell system.

Dcr-1 was cotransfected with the PA loqs isoform, the PB loqs isoform, or both simultaneously.

Dcr-1 can interact with either the PA or PB isoforms of loqs, but preferentially binds the PB isoform in competition experiments.

Source was cell extract of baculovirus-infected Sf21 cell line; bait produced from transfected construct; prey produced from transfected construct.

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

Residues 338-384 of the loqs PB isoform (which are not present in the PA isoform) increase the affinity for Dcr-1.

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

Dcr-2 and Dcr-1 preferentially interact with different isoforms of loqs.

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

Dcr-2 and Dcr-1 preferentially interact with different loqs isoforms.

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced as a recombinant fusion protein in Sf21 baculovirus-infected insect cell system.

The dsRBD3 domain of loqs is sufficient for only weak (~10%) binding of Dcr-1. The linker (L2 loop) of loqs is required in addition to the dsRBD3 domain for full binding to Dcr-1.

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced as a recombinant fusion protein in Sf21 baculovirus-infected insect cell system.

Deletion of the loqs linker (L2 loop) reduces binding to Dcr-1 ten-fold.

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

The addition of pre-miRNA resulted in an open conformation of the Dcr-1 interaction with loqs.