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Structural studies on the nucleocaps...

Green, Todd Jason.

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Structural studies on the nucleocapsid protein of vesicular stomatitis virus.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Structural studies on the nucleocapsid protein of vesicular stomatitis virus./
Author:	Green, Todd Jason.
Description:	156 p.
Notes:	Source: Dissertation Abstracts International, Volume: 63-10, Section: B, page: 4484.
Contained By:	Dissertation Abstracts International63-10B.
Subject:	Biology, Microbiology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3066312
ISBN:	0493856021

Structural studies on the nucleocapsid protein of vesicular stomatitis virus.
Green, Todd Jason.

Structural studies on the nucleocapsid protein of vesicular stomatitis virus. - 156 p.

Source: Dissertation Abstracts International, Volume: 63-10, Section: B, page: 4484.

Thesis (Ph.D.)--The University of Alabama at Birmingham, 2002.

This dissertation reports biological and structural studies on the nucleocapsid (N) protein from vesicular stomatitis virus (VSV). VSV is a nonsegmented, negative-stranded RNA virus belonging to the rhabdovirus family. The N protein is a multi-functional protein with functions that include (1) encapsidation of the viral genome with the goals of protecting the genetic material, in addition to presentation of this genetic material to the viral polymerase for the processes of transcription and replication; and (2) giving structure to the virus. The viral phosphoprotein (P protein) is responsible for complexing the N protein prior to performing its function of encapsidating VSV genomic RNA, thus preventing the concentration-dependent aggregation of the N protein. To derive structural information about the N protein and also to obtain soluble protein, the N protein and the P protein were expressed together in Escherichia coli. When coexpressed, the N and P proteins formed soluble protein complexes of various molar ratios. The major N/P protein complex was composed of 10 molecules of the N protein, 5 molecules of the P protein, and an RNA. The bacterially expressed N protein was determined to be functional through the binding activities with both the P protein and RNA. It was shown that N protein binding was localized to the C-terminal 72 amino acids of the P protein. This region of the P protein not only was sufficient for soluble N protein production but also proved to be adequate for preparing the N protein for RNA encapsidation. From NIP protein-RNA complexes, a single N protein-RNA complex was isolated. This N protein-RNA complex, devoid of the P protein, has been studied by several biochemical and physical methods, including analytical ultracentrifugation, electron microscopy, and X-ray crystallography. The N protein-RNA oligomer has been crystallized, and diffraction data has been collected. Results from the electron microscopy and crystallography experiments have shown that the N protein molecules are arranged with a 10-fold rotational symmetry giving the oligomer a disk-like morphology. The orientation of the N protein-RNA particle in the crystals has been investigated by self- and cross-rotation function calculations and translation searches by R-factor. The process for determining the high resolution structure of this complex is still in progress. However, a low resolution model of the N protein-RNA complex has been determined by image reconstruction techniques using negative-stain electron microscopy images.

ISBN: 0493856021Subjects--Topical Terms:

1017734
Biology, Microbiology.

Structural studies on the nucleocapsid protein of vesicular stomatitis virus.
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500 $a Source: Dissertation Abstracts International, Volume: 63-10, Section: B, page: 4484.
500 $a Chair: Ming Luo.
502 $a Thesis (Ph.D.)--The University of Alabama at Birmingham, 2002.
520 $a This dissertation reports biological and structural studies on the nucleocapsid (N) protein from vesicular stomatitis virus (VSV). VSV is a nonsegmented, negative-stranded RNA virus belonging to the rhabdovirus family. The N protein is a multi-functional protein with functions that include (1) encapsidation of the viral genome with the goals of protecting the genetic material, in addition to presentation of this genetic material to the viral polymerase for the processes of transcription and replication; and (2) giving structure to the virus. The viral phosphoprotein (P protein) is responsible for complexing the N protein prior to performing its function of encapsidating VSV genomic RNA, thus preventing the concentration-dependent aggregation of the N protein. To derive structural information about the N protein and also to obtain soluble protein, the N protein and the P protein were expressed together in Escherichia coli. When coexpressed, the N and P proteins formed soluble protein complexes of various molar ratios. The major N/P protein complex was composed of 10 molecules of the N protein, 5 molecules of the P protein, and an RNA. The bacterially expressed N protein was determined to be functional through the binding activities with both the P protein and RNA. It was shown that N protein binding was localized to the C-terminal 72 amino acids of the P protein. This region of the P protein not only was sufficient for soluble N protein production but also proved to be adequate for preparing the N protein for RNA encapsidation. From NIP protein-RNA complexes, a single N protein-RNA complex was isolated. This N protein-RNA complex, devoid of the P protein, has been studied by several biochemical and physical methods, including analytical ultracentrifugation, electron microscopy, and X-ray crystallography. The N protein-RNA oligomer has been crystallized, and diffraction data has been collected. Results from the electron microscopy and crystallography experiments have shown that the N protein molecules are arranged with a 10-fold rotational symmetry giving the oligomer a disk-like morphology. The orientation of the N protein-RNA particle in the crystals has been investigated by self- and cross-rotation function calculations and translation searches by R-factor. The process for determining the high resolution structure of this complex is still in progress. However, a low resolution model of the N protein-RNA complex has been determined by image reconstruction techniques using negative-stain electron microscopy images.
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