Poster Title

Determination of Domain Requirements of EF-G for Ribosome-Dependent GTPase Activity

Co-Author(s)

Catherine Shelton

Research Mentor(s)

Clint Spiegel

Affiliated Department

Chemistry

Sort Order

10

Start Date

14-5-2015 10:00 AM

End Date

14-5-2015 2:00 PM

Document Type

Event

Abstract

Requisite for all forms of life, the ribosome is a highly conserved, extremely complex ribonucleoprotein that functions to translate an organism's genetic information into their corresponding proteins. During this translation the two large subunits of the ribosome (30S and 50S) rotate with respect to one another, often forming a 'hybrid state', aided by a vast array of translation factors. Elongation-factor G (EF-G), a GTPase, catalyzes the translocation of the tRNA and mRNA through the ribosome after the addition of an amino acid to a polypeptide through the hydrolysis of GTP. Several domain mutants of EF-G were designed and purified, and the GTPase activity of each was tested, relative to the full length protein. EF-GΔ5 demonstrates a slight decrease in GTP hydrolysis relative to EF-G, maintaining ~85%. EF-GΔ4 and EF-GΔ4,5 maintain similar levels of activity, roughly 65% that of endogenous EF-G. EF-GΔG’ exhibits very low activity, ~10%, indicating this domain is very important in GTP hydrolysis.

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May 14th, 10:00 AM May 14th, 2:00 PM

Determination of Domain Requirements of EF-G for Ribosome-Dependent GTPase Activity

Chemistry

Requisite for all forms of life, the ribosome is a highly conserved, extremely complex ribonucleoprotein that functions to translate an organism's genetic information into their corresponding proteins. During this translation the two large subunits of the ribosome (30S and 50S) rotate with respect to one another, often forming a 'hybrid state', aided by a vast array of translation factors. Elongation-factor G (EF-G), a GTPase, catalyzes the translocation of the tRNA and mRNA through the ribosome after the addition of an amino acid to a polypeptide through the hydrolysis of GTP. Several domain mutants of EF-G were designed and purified, and the GTPase activity of each was tested, relative to the full length protein. EF-GΔ5 demonstrates a slight decrease in GTP hydrolysis relative to EF-G, maintaining ~85%. EF-GΔ4 and EF-GΔ4,5 maintain similar levels of activity, roughly 65% that of endogenous EF-G. EF-GΔG’ exhibits very low activity, ~10%, indicating this domain is very important in GTP hydrolysis.