Tethered GLD-2 family members. (A) Protocol. mRNA encoding an MS2 fusion protein with the candidate polypeptide was injected into oocytes. The fusion protein was expressed during a 6-h incubation, and then two reporter mRNAs were coinjected. The luciferase mRNA contained MS2 binding sites; the β-galactosidase lacked MS2 sites, and was used as a control. (B) Results. (Top) Relative stimulation of translation by each protein, expressed as the ratio of luciferase to β-galactosidase activities. Values were normalized to CeGLD-2 D608A, a catalytically inactive form of the enzyme (Kwak et al. 2004). Error bars were derived from more than three experiments. (Bottom) Chimeric fusion proteins detected by Western blotting using anti-HA tag antibody. Lysates equivalent to three oocytes were loaded into each lane.
