SPMS MS02 02 Sin ingle le-Mo Molec lecule ule Bi Biophys - PowerPoint PPT Presentation

SPMS MS02 02 Sin ingle le-Mo Molec lecule ule Bi Biophys hysics: ics: Mec Mecha hani nica cal l Un Unfo fold ldin ing g of R f RNA NA by y Optica ical l Tw Twee eezer ers

In Introduc oducti tion on Objective: Investigate a potential RNA triplex structure stimulating ribosomal frameshifting through Mechanical pulling of target RNA in different chemical environments using optical tweezer

Tr Trip iplex lex RNA NA serves catalytic and regulatory functions in cellular important nucleoprotein structural motif complexes found in pseudoknots and structured RNA enables recoding of forms through ribosomal tertiary frameshifting interactions in the major or minor groove of a Watson-Crick base-paired stem

Rib ibos osomal omal Fr Fram ameshift eshifting ing

Op Opti tical al Tw Twee eezer er

Bu Buffers ffers Us Used ed (i) 200 mM NaCl, 20 mM NaOAc, 0.1 mM EDTA, pH 5.0 and (ii) 5 mM MgCl2 , 100 μM spermine , 200 mM NaCl, 20 mM NaOAc, 0.1 mM EDTA, pH 5.0. lowered pH is in favour of the formation of the triplex structure

Pu Pull llin ing g Mod odes es Pulling Modes Force-ramp Constant-force

Pu Pull llin ing g Mod ode: e: Fo Force-ramp amp

Pu Pull llin ing g Mod ode: e: Co Constan tant-force force Diagram of raw data for forces 16.5pN, 16.8pN, 17.1pN and 17.4pN in buffer 1 Diagram of raw data for forces 17.4pN, 17.7pN, 18.0pN and 18.3pN in buffer 2

Cr Crit itic ical al Fo Force Buffer 1: Critical force= 17.0pN Buffer 2: Critical force= 18.1pN

What at I h I hav ave e le lear arnt The role of triplex in stabilising RNA The correlation Spermine between and Mg 2+ mechanical stabilize the property and RNA stability of RNA structure. triplex

Im Impr provement ovement Melting studies done by Ultraviolet absorption Melting temperature (Tm) Thermodynamic information (stability) on triplex RNA

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