An Investigation of Intersubunit Bridges Important for Ribosome Stability in Saccharomyces Cerevisiae
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Author
Ackell, Andrew Peter Hummel
Subject
Washington and Lee University -- Honors in Chemistry
Ribosomes
Proteins -- Stability
Saccharomyces cerevisiae
Metadata
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The ribosome, a large ribonucleoprotein complex responsible for protein
biosynthesis, is composed of two unequal subunits that must correctly associate for
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proper function. X-ray crystallographic and cryo-EM studies have identified a number of
specific interactions, or intersubunit bridges (ISBs ), between the large and small subunits.
At least two of these ISBs have been shown to be important for ribosome stability in
bacteria using a technique known as modification interference. A similar modification
interference approach is presented to identify ISBs that are important for ribosome
stability in yeast. Native ribosomes and ribosomal subunits were isolated from yeast cells.
Sucrose density gradient ultracentrifugation under a variety of conditions was
investigated in order to achieve separation and purification of individual ribosomal
subunits from intact ribosomes. The determination of the optimal conditions for subunit
separation and isolation is a necessary step toward identifying possible ISBs through
modification int~rf erence. We hope our modification interference investigations will
provide insights regarding the function and conservation of ISBs.