Ribosome collisions alter frameshifting at translational reprogramming motifs in bacterial mRNAs.
Translational frameshifting involves the repositioning of ribosomes on their messages into decoding frames that differ from those dictated during initiation. Some messenger RNAs (mRNAs) contain motifs that promote deliberate frameshifting to regulate production of the encoded proteins. The mechanisms of frameshifting have been investigated in many systems, and the resulting models generally involve single ribosomes responding to stimulator sequences in their engaged mRNAs. We discovered that the abundance of ribosomes on messages containing the IS3, dnaX, and prfB frameshift motifs significantly influences the levels of frameshifting. We show that this phenomenon results from ribosome collisions that occur during translational stalling, which can alter frameshifting in both the stalled and trailing ribosomes. Bacteria missing ribosomal protein bL9 are known to exhibit a reduction in reading frame maintenance and to have a strong dependence on elongation factor P (EFP). We discovered that ribosomes lacking bL9 become compacted closer together during collisions and that the E-sites of the stalled ribosomes appear to become blocked, which suggests subsequent transpeptidation in transiently stalled ribosomes may become compromised in the absence of bL9. In addition, we determined that bL9 can suppress frameshifting of its host ribosome, likely by regulating E-site dynamics. These findings provide mechanistic insight into the behavior of colliding ribosomes during translation and suggest naturally occurring frameshift elements may be regulated by the abundance of ribosomes relative to an mRNA pool.
['Smith|Angela M|AM|', 'Costello|Michael S|MS|', 'Kettring|Andrew H|AH|', 'Wingo|Robert J|RJ|', 'Moore|Sean D|SD|0000-0003-1606-7771']
D004926:Escherichia coli D016368:Frameshift Mutation D018965:Frameshifting, Ribosomal D009690:Nucleic Acid Conformation D010445:Peptide Elongation Factors D014176:Protein Biosynthesis D012333:RNA, Messenger D016364:Reading Frames D012269:Ribosomal Proteins D012270:Ribosomes
ribosome frameshift translation dnaX bL9