FIGURE 4.

Ty3 VLPs associate with P-body proteins. (A) Ty3-RFP colocalizes with P-body proteins fused to GFP. Yeast strain BY4741 (wild type) and strains derived from it by fusion of the GFP coding region in frame downstream of genes encoding P-body proteins (labeled Xrn1, etc.) and for the gene encoding Nvj1, a nuclear-vacuolar junction protein, were transformed with the Ty3-RFP or RFP expression plasmids and grown in the presence of galactose for 4 h. No Ty3 refers to the strain containing RFP expressed from the galactose-inducible promoter in pYES2.0. RFP alone expressed at high levels does not form foci, and Nvj1-GFP does not colocalize with Ty3-RFP. Unmarked cells (wild type) do not show bleed-through of Ty3-RFP into the green channel. Scale bar, 5.0 μm. (B) Dhh1-GFP and Xrn1/Kem1-GFP are associated with Ty3 VLP clusters. Cells induced for expression of Ty3 for 4 h were processed for EM and reacted with anti-GFP and secondary antibodies conjugated to colloidal gold. Solid arrows indicate colloidal gold particles. Scale bar, 0.1 μm.