Single-molecule tracking reveals dynamic regulation of ribosomal scanning.

Hong, Hea Jin; Zhang, Antonia L; Conn, Adam B; Blaha, Gregor; O'Leary, Seán E · Sci Adv · 2024

basic_science · Level V

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Abstract

How eukaryotic ribosomes traverse messenger RNA (mRNA) leader sequences to search for protein-synthesis start sites remains one of the most mysterious aspects of translation and its regulation. While the search process is conventionally described by a linear "scanning" model, its exquisitely dynamic nature has restricted detailed mechanistic study. Here, we observed single <i>Saccharomyces cerevisiae</i> ribosomal scanning complexes in real time, finding that they scan diverse mRNA leaders at a rate of 10 to 20 nt s<sup>-1</sup>. We show that specific binding of a protein to its mRNA leader sequence substantially arrests scanning. Conversely, impairing scanning-complex guanosine 5'-triphosphate hydrolysis results in native start-site bypass. Our results illustrate an mRNA-centric, kinetically controlled regulatory model where the ribosomal pre-initiation complex amplifies a nuanced energetic landscape to regulate scanning and start-site selection fidelity.

Medical subject headings