Abstract Title

Investigation of the minimal binding substrate for RNA modification enzyme RsuA

Abstract

Ribosomes are important for all living organisms. Nucleotide modification found in various regions of the ribosomal RNA can influence ribosomal assembly, local structural, and thermodynamic stability changes in rRNA. At the same time modification enzymes can influence the binding thermodynamics and kinetics of various ribosomal proteins, hence influencing ribosome assembly. In particular, pseudouridine synthase, RsuA, is responsible for modifying uridine at position 516 in the 16S rRNA to a pseudouridine that can influence the structure and thermodynamic stability of 16S helix 18. Previous studies have shown that protein RsuA is unable to bind to 16S rRNA alone and can only weakly bind to 30S ribosomes. It is proposed that protein RsuA is binding to a ribosome assembly intermediate. However, this assembly intermediate is yet be discovered. We have successfully overexpressed, purified and fluorescently labeled the RsuA protein. We are currently investigating the binding thermodynamics of RsuA enzyme in the presence of various ribosomal RNA substrates to determine the high affinity substrate for protein RsuA.

Modified Abstract

Ribosomes are important for all living organisms. Nucleotide modification found in various regions of the ribosomal RNA can influence ribosomal assembly, local structural, and thermodynamic stability changes. At the same time modification enzymes can influence the binding thermodynamics and kinetics of various ribosomal proteins, hence influencing ribosome assembly. In particular, pseudouridine synthase, RsuA, is responsible for modifying uridine at position 516 in the 16S rRNA to a pseudouridine that can influence the structure and thermodynamic stability of 16S helix 18. We have successfully overexpressed, purified and fluorescently labeled the RsuA protein. We are currently investigating the binding thermodynamics of RsuA enzyme in the presence of various ribosomal RNA substrates to determine the high affinity substrate for protein RsuA.

Research Category

Biology/Ecology

Primary Author's Major

Biochemistry

Mentor #1 Information

Kumudie Jayalath

Mentor #2 Information

Dr. Sanjaya Abeysirigunawardena

Presentation Format

Poster

Start Date

March 2016

Research Area

Biochemistry | Biophysics

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Mar 15th, 1:00 PM

Investigation of the minimal binding substrate for RNA modification enzyme RsuA

Ribosomes are important for all living organisms. Nucleotide modification found in various regions of the ribosomal RNA can influence ribosomal assembly, local structural, and thermodynamic stability changes in rRNA. At the same time modification enzymes can influence the binding thermodynamics and kinetics of various ribosomal proteins, hence influencing ribosome assembly. In particular, pseudouridine synthase, RsuA, is responsible for modifying uridine at position 516 in the 16S rRNA to a pseudouridine that can influence the structure and thermodynamic stability of 16S helix 18. Previous studies have shown that protein RsuA is unable to bind to 16S rRNA alone and can only weakly bind to 30S ribosomes. It is proposed that protein RsuA is binding to a ribosome assembly intermediate. However, this assembly intermediate is yet be discovered. We have successfully overexpressed, purified and fluorescently labeled the RsuA protein. We are currently investigating the binding thermodynamics of RsuA enzyme in the presence of various ribosomal RNA substrates to determine the high affinity substrate for protein RsuA.