121st General Meeting of the KCS

Type Poster Presentation
Area Life Chemistry
Room No. Event Hall
Time 4월 20일 (금요일) 11:00~12:30
Code BIO.P-282
Subject Investigation of the tertiary interaction U4/U6 snRNP recycling factor and U6 snRNA
Authors Kyeong-Mi Bang, Ji-Yeon Shin, Nak-Kyoon Kim1,*
Korea Institute of Science and Technology, Korea
1Advanced Analysis Center, Korea Institute of Science and Technology, Korea
Abstract Splicing is a catalytic process, in which pre-mRNAs are converted to mature mRNAs by the spliceosome. As a result of splicing, the two exons are connected and an intron lariat is released. Spliceosome is a large and dynamic RNA-protein complex, which is made up of five small nuclear ribonucleoproteins (U1, U2, U4, U5, and U6 snRNPs). In the splicing process, the components of spliceosome are recycled for a new round of catalysis. Among many recycling factors, squamous cell carcinoma antigen recognized by T-cells 3 (SART3) is required to adopt the U6 snRNA for the formation of the U4/U6 snRNP complex in human. SART3 consists of a 12 half-a-tetratricopeptide (HAT) repeat domains in N-terminal, a nuclear localization signal (NLS) sequence, two RNA recognition motifs (RRMs), and an extension of 10 highly conserved amino acids in C-terminal (C10). Among these domains, the RRM domains are suggested to be involved in the binding of the U6 snRNA. U6 snRNA is believed to play an essential role in the catalytic mechanism of pre-mRNA splicing, and the highly conserved 6 nucleotides, ACAGAG, present in the U6 snRNA are especially associated with the interaction of SART3 RRM domains. In this study, I designed several constructs of U6 snRNA containing ACAGAG, which are predicted to interact with RRMs of SART3, and confirmed the tertiary interaction between RRMs of SART3 and U6 snRNA using electrophoretic mobility shift assay (EMSA). Also, I performed a structural investigation of the tertiary interaction between RRMs and U6 snRNA using NMR spectroscopy. These results will provide a structural basis for the complex formation of U4/U6 snRNP mediated by SART3.
E-mail bangkmi9@gmail.com