Figure 4. Ribosome footprinting-based methods for genome-wide translatome surveys.

a. Ribosome profiling is based on deep sequencing of RNA fragment protected by the ribosome during RNase digestion.

b. Ribosome footprints from translation initiation sites are highly enriched by the use of translation inhibitors LTM or harringtonine.

c. Artificially localized BirA (ex. on the ER or mitochondria) specifically biotinylates Avi-tags on ribosome proteins, only when those two molecules are coincidentally localized. Purification and footprinting of biotinylated ribosomes thereby surveys the translatome at a specific place within cells.

d. Co-translational events can be monitored transcriptome-wide by profiling of ribosomes selected through interacting factors.

e. In TCP-Seq (translation complex profiling sequencing), the scanning 40S ribosome and its footprint are crosslinked, purified, and sequenced.