Fig. 3.
Inhibited BMV RNA replication in bro1Δ cells correlates with a lack of Doa4p's enzymatic activity and a decrease in free Ub levels. (A) Inhibited BMV RNA replication is restored by supplementation with wt Bro1p in bro1Δ cells. (B) BMV RNA replication is related to the levels of free Ub in bro1Δ cells. The bro1-2 mutant was expressed from either a low- or high-copy-number plasmid, and mutant bro1-C was expressed from a low-copy-number plasmid in bro1Δ cells. Note that the anti-Bro1p antiserum did not detect bro1-C. comp., complementing. (C) The functional interaction between the C termini of Doa4p and Bro1p is required for BMV RNA replication. The doa4AAFA mutant, which does not interact with Bro1p or support cargo sorting and deubiquitination, did not restore BMV RNA replication in doa4Δ cells. Accumulation of BMV RNAs and Ub, Bro1p, and Pgk1p levels were detected as in Fig. 1.