Determination of Domain Requirements of EF-G for Ribosome-Dependent GTPase Activity
Research Mentor(s)
Spiegel, P. Clint
Description
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.
Document Type
Event
Start Date
14-5-2015 10:00 AM
End Date
14-5-2015 2:00 PM
Department
Chemistry
Genre/Form
student projects; posters
Subjects – Topical (LCSH)
GTPase-activating protein; Ribosomes
Type
Image
Rights
Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this documentation for commercial purposes, or for financial gain, shall not be allowed without the author's written permission.
Language
English
Format
application/pdf
Determination of Domain Requirements of EF-G for Ribosome-Dependent GTPase Activity
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.