In vivo targets and mechanisms of gene control during protein synthesis
Dr Nikolay Shirokikh, The Preiss Group- RNA Biology, JCSMR
Translation of messenger(m)RNA into proteins is central to eukaryotic gene control. Dysregulated translation is common in disease, including cancer. Mechanisms defining mRNA-specific rates of protein synthesis are key to understanding translational regulation, yet their identification was hampered for decades by the inability to trap and isolate most of the in vivo translation intermediates.
To overcome these limitations, we re-considered high-throughput approaches to protein synthesis and developed translation complex profile sequencing (TCP-seq). Using TCP-seq, we can capture translation intermediates of all types as they natively occur in vivo and answer long-standing mechanistic questions of translation in yeast. We further provide mRNA-specific information on the extent of control and possibilities to directly identify types and locations of the responsible RNA regulatory elements. We are now aiming to use TCP-seq for mechanistic discovery in mammalian cells and for the analysis of translational dysregulation in tumours to identify novel druggable targets.