Fig. 1: Chp1 binds ribosomes aided by direct interaction with the α-subunit of NAC.

a Hsf1 activity in yeast strains with single deletions of genes encoding either ribosome-associated proteins (RAP, blue) or non-ribosome associated proteins (NRAP, orange). b Analysis of functional categories of Chp1-interacting proteins identified by in vivo site-specific UV crosslinking followed by two-step affinity purification and peptide mass fingerprinting identification using LC-MS. c Chp1-GFP and NAC subunits co-IP from the indicated yeast strains. The experiment was performed three times. d Western blot analysis of Ni-NTA matrix binding of Chp1-Myc-6His mixed with NAC (Egd1- Egd2) or NAC lacking the UBA domain (Egd1- Egd2 ΔUBA). The experiment was performed three times. e Total soluble yeast cell lysate (T) was separated into ribosomal (R) and soluble (S) fractions in the presence of either 50 mM or 500 mM KOAc and the localization of Chp1-GFP, Egd2 and Rpl25 among the different fractions was analyzed by western blot. The experiment was performed three times. f Chp1-GFP and Rpl2A co-IP from the indicated yeast strains. The experiment was performed three times.