Title:
Structural studies of ribonucleoprotein complexes using molecular modeling
Structural studies of ribonucleoprotein complexes using molecular modeling
Author(s)
Devkota, Batsal
Advisor(s)
Harvey, Stephen C.
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Abstract
The current work reports on structural studies of ribonucleoprotein complexes, Escherichia coli and Thermomyces lanuginosus ribosomes, and Pariacoto virus (PaV) using molecular modeling. Molecular modeling is the integration and representation of the structural data of molecules as models. Integrating high-resolution crystal structures available for the E. coli ribosome and the cryo-EM density maps for the PRE- and POST- accommodation states of the translational cycle, I generated two all-atom models for the ribosome in two functional states of the cycle. A program for flexible fitting of the crystal structures into low-resolution maps, YUP.scx, was used to generate the models. Based on these models, we hypothesize that the kinking of the tRNA plays a major role in cognate tRNA selection during accommodation.
Secondly, we proposed all-atom models for the eukaryotic ribosomal RNA. This is part of a collaboration between Joachim Frank, Andrej Sali, and our lab to generate an all-atom model for the eukaryotic ribosome based on a cryo-EM density map of T. lanuginosus available at 8.9Å resolution. Homology modeling and ab initio RNA modeling were used to generate the rRNA components.
Finally, we propose a first-order model for a T=3, icosahedral, RNA virus called Pariacoto virus. We used the structure available from x-ray crystallography as the starting model and modeled all the unresolved RNA and protein residues. Only 35% of the total RNA genome and 88% of the protein were resolved in the crystal structure. The generated models for the virus helped us determine the location of the missing N-terminal protein tails. The models were used to propose a new viral assembly pathway for small RNA viruses. We propose that the basic N-terminal tails make contact with the RNA genome and neutralize the negative charges in RNA and subsequently collapse the RNA/protein complex into a mature virus. This process is reminiscent of DNA condensation by positively charged ions.
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Date Issued
2007-12-06
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Text
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Dissertation