Fig. 3: Collided disomes accumulate in cells expressing catalytically inactive Hel2-RM or lacking the 40S ribosomal protein Asc1.
a The bulk of Hel2-RM binds to disomes and trisomes Polysome profiles with extracts derived from Hel2-RMg-log (orange) or Hel2-RMg-log/RNase (magenta). b Hel2-RM binds to ribosomes in a salt-resistant manner. Ribosome binding assays with extract of wild typelog or Hel2-RMlog as described in Fig. 2c. Shown is the mean of 3 independent experiments (bars) and the result of each experiment (dots) with the standard deviation. An example blot is shown in Fig. S3a. c High affinity binding of Hel2 to disomes depends on Asc1. Polysome profiles with extracts of ∆asc1log (orange) or ∆asc1log/RNase (magenta). d Hel2 binds to ∆asc1-disomes in a salt-sensitive manner, with or without induction of collision. Ribosome-binding assays were performed with extract of ∆asc1log (no-collision, blue) or ∆asc1coll (collision, light blue). Shown is the mean of at least 3 independent experiments (bars) and the result of each experiment (dots). An example blot is shown in Fig. S3d. Polysome profiles shown in a and c, including wild typelog/RNase A260 traces (dotted gray lines) and Hel2 distribution profiles of wild typelog/RNase (gray), were recorded side by side. Labeling is as detailed in Fig. 1.
