[대학원 생명과학과 세미나 안내]
연사 : 김종서 교수(서울대학교, 생명과학부)
연제 : System-wide Mapping of RNA-Protein Interactions via Mass Spectrometry
일시 : 2021년 03월 12일 (금) 오후 5시
장소 : 온라인 화상 강의로 진행됩니다.
초청교수 : 우재성 교수
Abstract
Mass spectrometry (MS) based proteomics tool can uniquely provide the information of the post-translational modifications and protein-to-protein interactions, compared to the NGS-based transcriptomics. Furthermore, recent studies reported largely low correlation between mRNA and protein expression levels, especially for lowly expressed proteins. In this regard, highly sensitive and unbiased proteomics platform is highly desirable for biological researches. In this presentation, I will introduce ultrasensitive LC-MS system to overcome the innate hurdle of MS-based proteomics in terms of sensitivity. And I will present a couple of studies solely enabled by my advanced LC-MS system along with my recent biochemical developments in RNA biology; 1) RBS-ID and 2) FAX-RIC.
RNA-binding site (RBS)-ID is a method that uses hydrofluoride to fully cleave RNA into mono-nucleosides, thereby minimizing the search space to drastically enhance coverage and to reach single amino acid resolution. The simple mono-nucleoside adducts offer a confident and quantitative measure of direct RNA–protein interaction. Using RBS-ID, we profiled ~2,000 human RBSs and probed Streptococcus pyogenes Cas9 to discover residues important for genome editing. I also introduce formaldehyde crosslinking (FAX) as an alternative chemical crosslinking for RNA interactome capture (RIC). Mild FAX captures RNA–protein interaction with high specificity and efficiency in cell culture. Furthermore, FAX-RIC is applicable to mammalian tissue samples such as Mus musculus liver, extending the RNA interactome profiling into multicellular organisms.