Live-cell single RNA imaging reveals bursts of translational frameshifting

Kenneth Lyon; Luis U. Aguilera; Tatsuya Morisaki; Brian Munsky; Timothy J. Stasevich

To measure the average elongation rate, harringtonine (Cayman Chemical) was used to block translation initiation and iduce the run-off of all actively elongating ribosomes, leading to a gradual loss of nascent chain signal at translation sites. Beadloaded cells with visible translation sites were imaged as described above except that cell volumes were acquired every 60 or 120 seconds. After acquiring 5 pre-images, cells were treated with a final concentration of 3 µg/ml Harringtonine and continuously imaged for 30 more time points. As a control, the same imaging conditions were performed except that cells were treated with vehicle (DMSO). In this case, nascent chain signals did not decay (data not shown). The intensities of translation sites measured, as described above. The All rights reserved. No reuse allowed without permission.