Pum
Halotag
AGO2
FLAG tag
coordinates relative to 1533 aa isoform
Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.
The pum T1137E mutation does not disrupt binding to AGO2.
No interaction between pum and AGO1 was detected in this assay.
Halotag pulldown, TEV elution and FLAG Western blot detection of prey.
V5 tag, MS2 fusion
Deletion of various PUF repeats in the pum RNA-binding domain (RBD) abrogated binding to AGO2.
Deletion of pum PUF repeats 7-8 abrogated binding to AGO2, but did not affect pum repressive activity in a reporter assay.
