Year: 2022
Type: article
Abstract: Cellular global translation is often measured using ribosome profiling or quantitative mass spectrometry, but these methods do not provide direct information at the level of elongating nascent polypeptide chains (NPCs) and associated co-translational events. Here, we describe pSNAP, a method for proteome-wide profiling of NPCs by affinity enrichment of puromycin- and stable isotope-labeled polypeptides. pSNAP does not require ribosome purification and/or chemical labeling, and captures bona fide NPCs that characteristically exhibit protein N-terminus-biased positions. We applied pSNAP to evaluate the effect of silmitasertib, a potential molecular therapy for cancer, and revealed acute translational repression through casein kinase II and mTOR pathways. We also characterized modifications on NPCs and demonstrated that the combination of different types of modifications, such as acetylation and phosphorylation in the N-terminal region of histone H1.5, can modulate interactions with ribosome-associated factors. Thus, pSNAP provides a framework for dissecting co-translational regulations on a proteome-wide scale.
Source: iScience
Institutions: Kyoto University, RIKEN Center for Biosystems Dynamics Research, Institute for Integrated Cell-Material Sciences, Kyoto University, Shenyang Pharmaceutical University, Center for iPS Cell Research and Application, Kyoto University, Kyoto Sangyo University, National Institute of Biomedical Innovation, Health and Nutrition, Japan Science and Technology Agency, RIKEN Center for Integrative Medical Sciences
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,000
Funders: Dow Chemical Company, Harvard Medical School, Takeda Science Foundation, Japan Agency for Medical Research and Development, Tokyo Institute of Technology, Japan Society for the Promotion of Science, Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology, Exploratory Research for Advanced Technology
OpenAlex ID: W4281963797