Pleiohomeotic Interacts with the Core Transcription Elongation Factor Spt5 to Regulate Gene Expression in Drosophila
Figure 1
Pho physically interacts with Spt5.
A) Yeast 2-hybid assay showing binding of full length Pho with the Spt5 DD domain and full length Spt5. Vector (pGBKT7) containing no insert was used as a control to demonstrate that Pho does not activate reporter gene expression in the absence of Spt5. B) Pho binds to immobilized GST-DD. Ten percent of the input Pho is run in left lane, immobilized GST in middle lane incubated with Pho as negative control. C) Western blots of co-immunoprecipitation (co-IP) assays from S2 cell extracts of Flag-tagged Spt5 with Myc-Spt4 (positive control), Myc-Pho, Myc-N-Pho (amino acids 1–351), Myc-C-Pho (351–520), Myc-GFP (negative control) and no protein. D) Western blots of co-IP assays from S2 cell extracts of Flag-tagged W049 variant of Spt5 with Myc-Spt4 (positive control), Myc-Pho, Myc-GFP (negative control) and no protein.