Yeast Bud27 modulates the biogenesis of Rpc128 and Rpc160 subunits and the assembly of RNA polymerase III

•Yeast Bud27 interacts with RNA polymerase (pol) III and affects its transcription.•Pol III requires Bud27 for interaction with RSC and Hsp70 family member Ssa4.•Nuclear import of the assembled pol III. requires Ssa4 and its nuclear exporter Msn5.•Bud27 affects the synthesis of Rpc160 and Rpc128 subunits of pol III differently.•The altered Rpc160/Rpc128 ratio leads to defective pol III assembly in the cytoplasm.

Yeast Bud27, an unconventional prefoldin is reported to affect the expression of nutrient-responsive genes, translation initiation and assembly of the multi-subunit eukaryotic RNA polymerases (pols), at a late step. We found that Bud27 associates with pol III in active as well as repressed states. Pol III transcription and occupancy at the target genes reduce with the deletion of BUD27. It promotes the interaction of pol III with the chromatin remodeler RSC found on most of the pol III targets, and with the heat shock protein Ssa4, which helps in nuclear import of the assembled pol III. Under nutrient-starvation, Ssa4–pol III interaction increases, while pol III remains inside the nucleus. Bud27 but not Ssa4 is required for RSC–pol III interaction, which reduces under nutrient-starvation. In the bud27Δ cells, total protein level of the largest pol III subunit Rpc160 but not of Rpc128, Rpc34 and Rpc53 subunits is reduced. This is accompanied by lower transcription of RPC128 gene and lower RPC160 translation due to reduced association of mRNA with the ribosomes. The resultant alteration in the normal cellular ratio of the two largest subunits of pol III core leads to reduced association of other pol III subunits and hampers the normal assembly of pol III at an early step in the cytoplasm. Our results show that Bud27 is required in multiple activities responsible for pol III biogenesis and activity.