Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey from ovary cDNA expression library).
Two-hybrid system: yeast LexA-BD/B42-AD
Dpit47 associates with about 10% of cellular DNApol-α180 at all stages of embryogenesis. Dpit47 also associates with DNApol-α60, DNApol-α73, but not with PCNA, Cdk2, Top2 or lamins.
The Dpit47-DNApol-α180 interaction is stabilized by 1 ug/ml geldanamycin, which binds the ATP-binding site of Hsp83 to block its chaperone activity. On the other hand, the addition of 1mM ATP, which catalyzes the release of Hsp83 substrates, destabilizes the Dpit47-DNApol-α180 interaction. This suggests that DNApol-α180 is a substrate for Hsp83 chaperone activity, with Dpit47 acting as a co-chaperone.
DNApol-α180 associated with Dpit47 is inactive in DNA polymerase assay, as compared to total DNApol-α180.
Source was embryos of wild-type fly line; bait produced from endogenous gene; prey produced from endogenous gene.
DNApol-α180 associates with Dpit47 in exponentially growing cells, but not in quiescent cells. This lack of interaction is not due to the absence of DNApol-α180 as it is detected at similar levels in quiescent cells.
Source was cell extract of S2 cell line; bait produced from endogenous gene; prey produced from endogenous gene.
