Fig 7.
Expressing CAP and HEL in trans failed to perform any full-length 1a functions. (A) Total protein was isolated from yeast expressing 1a or CAP plus HEL and analyzed by Western blotting using anti-1a antibodies. (B) Analysis of positive-strand RNA3 and RNA4 from yeast expressing 2apol, RNA3, and either wt 1a or a combination of the 1a fragments. Each set of conditions was tested in triplicate. (C) RNA3 levels in yeast expressing RNA3 alone or with wt 1a or with the 1a fragments. For panels B and C, total RNA was isolated from yeast cells and Northern blotting was used to examine the accumulation of RNA3 and RNA4 or RNA3 with a BMV RNA-specific probe. 18S rRNA was measured as a loading control. (D) EM of yeast cell coexpressing CAP and HEL. Bar, 200 nm. (E) Localization of GFP-tagged CAP and HA-tagged HEL expressed in trans. DNA was stained with DAPI (blue). Bar, 2 μm.