Interaction in vitro; bait produced and labeled by in vitro transcription; prey derived from wild-type adult female ovary extract.

The pum protein is detected as a 165 kDa band (close to its predicted 157.5 kDa size). Binding is absent in pum mutant extracts, and inhibited by pum antiserum.

Binding to the hb NRE by another unidentified 55 kDa factor (not nos or pum was observed).

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

Three-hybrid system: yeast LexA-BD/MS2-SL/GAL4-AD. LexA-BD fused to phage MS2 coat protein, which binds MS2-SL.

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

Three-hybrid system: yeast GAL4-BD/MS2-SL/GAL4-AD. GAL4-AD fused to phage MS2 coat protein, which binds MS2-SL.

Interaction in vitro; bait produced as a recombinant fusion proteins in bacterial system; prey produced and labeled in vitro.

Positive control.

Interaction in vitro; bait produced as a recombinant fusion proteins in bacterial system; prey produced and labeled in vitro.

Positive control.

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey oligonucleotide produced and labeled in vitro.

Cooperative binding of pum to hb NRE RNA element is best fit by a two-site model. Each hb NRE RNA element is composed of two pum binding sites: BoxA and BoxB.

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey oligonucleotide produced and labeled in vitro.

Data are consistent with binding or one or two molecules of pum protein to each hb NRE RNA element. Two pum-NRE complexes are observed in gels - the higher molecular weight complex is not observed when one of the two NRE boxes is mutated.

yeast LexA/GAL4-AD.

pum binds 3\' UTR NRE consensus sequences (UGUANAUA) in hb, Egfr, rl, Sos and drk, but not in phl (also known as raf).

Interaction in vitro; bait produced and labeled by in vitro transcription; prey produced and labeled by in vitro translation.

Positive control.

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro transcription.

Binding of pum to hb mRNA facilitated by prebinding mRNA with brat.

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

Ternary complex of nos and pum proteins and hb RNA.

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

Binding of hb NRE2 by pum protein is enhanced by nos protein. nos alone does not bind hb in this assay.

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

Positive control.