Thesis (M.S.)--Ohio State University. / Bibliography: leaves 108-109. Available online via OhioLINK's ETD Center

Ebola virus (EBOV) is an enveloped, negative-sense single-stranded RNA virus that causes severe hemorrhagic fever in humans and nonhuman primates. The EBOV glycoprotein (GP) gene encodes multiple transcripts due to RNA editing at a conserved editing site (ES) (a hepta-uridine stretche). The majority of GP gene transcript is unedited and encodes for a soluble glycoprotein (sGP); a defined function has not been assigned for sGP. In contrast, the transmembrane glycoprotein (GP1,2) dictates viral tropism and is expressed through RNA editing by insertion of a nontemplate adenosine (A) residue. Hypothetically, the insertion/deletion of a different number of A residues through RNA editing would result in another yet unidentified GP gene product, the small soluble glycoprotein (ssGP). I have shown that ssGP specific transcripts were indeed produced during EBOV infection. Detection of ssGP during infection was challenging due to the abundance of sGP over ssGP and the absence of distinguishing antibodies for ssGP. Optimized two- dimensional (2-D) gel electrophoresis verified the expression of ssGP during infection. Biophysical characterization revealed ssGP is a disulfide-linked homodimer that is exclusively N-glycosylated. Although ssGP appears to share similar structural properties with sGP, it does not have the same anti-inflammatory function. Using a new rapid transcript quantification assay (RTQA), I was able to demonstrate that RNA editing is an inherent feature of the genus Ebolavirus and all species of EBOV produce multiple GP gene products. A newly developed dual-reporter minigenome system was utilized to characterize EBOV RNA editing and determined the conserved ES sequence and cis-acting sequences as primary and secondary requirements for RNA editing, respectively. Viral protein (VP) 30, a transcription activator, was identified as a contributing factor of RNA editing— a proposed novel function for this largely uncharacterized viral protein. Finally, I could show that EBOV RNA editing is GP gene-specific because a similar sequence located in L gene did not serve as an ES, most likely due to the lack of the necessary cis-acting sequences. In conclusion, I identified a novel soluble protein of EBOV whose function needs further characterization. I also shed light into the mechanism of EBOV RNA editing, a potential novel target for intervention.

No description available.

Ebola virus (EBOV) is an enveloped, negative-sense single-stranded RNA virus that causes severe hemorrhagic fever in humans and nonhuman primates. The EBOV glycoprotein (GP) gene encodes multiple transcripts due to RNA editing at a conserved editing site (ES) (a hepta-uridine stretche). The majority of GP gene transcript is unedited and encodes for a soluble glycoprotein (sGP); a defined function has not been assigned for sGP. In contrast, the transmembrane glycoprotein (GP1,2) dictates viral tropism and is expressed through RNA editing by insertion of a nontemplate adenosine (A) residue. Hypothetically, the insertion/deletion of a different number of A residues through RNA editing would result in another yet unidentified GP gene product, the small soluble glycoprotein (ssGP). I have shown that ssGP specific transcripts were indeed produced during EBOV infection. Detection of ssGP during infection was challenging due to the abundance of sGP over ssGP and the absence of distinguishing antibodies for ssGP. Optimized two- dimensional (2-D) gel electrophoresis verified the expression of ssGP during infection. Biophysical characterization revealed ssGP is a disulfide-linked homodimer that is exclusively N-glycosylated. Although ssGP appears to share similar structural properties with sGP, it does not have the same anti-inflammatory function. Using a new rapid transcript quantification assay (RTQA), I was able to demonstrate that RNA editing is an inherent feature of the genus Ebolavirus and all species of EBOV produce multiple GP gene products. A newly developed dual-reporter minigenome system was utilized to characterize EBOV RNA editing and determined the conserved ES sequence and cis-acting sequences as primary and secondary requirements for RNA editing, respectively. Viral protein (VP) 30, a transcription activator, was identified as a contributing factor of RNA editing— a proposed novel function for this largely uncharacterized viral protein. Finally, I could show that EBOV RNA editing is GP gene-specific because a similar sequence located in L gene did not serve as an ES, most likely due to the lack of the necessary cis-acting sequences. In conclusion, I identified a novel soluble protein of EBOV whose function needs further characterization. I also shed light into the mechanism of EBOV RNA editing, a potential novel target for intervention.

Thesis (Ph. D.)--Colorado State University, 2004. / Includes bibliographical references.

Thesis, PlanB (M.S.)--University of Wisconsin--Stout, 1999. / Includes bibliographical references.

Thesis (M.S.)--Villanova University, 2010. / Chemistry Dept. Includes bibliographical references.

Thesis (Ph. D. in Pharmacology)--Vanderbilt University, Dec. 2005. / Title from title screen. Includes bibliographical references.

Thesis (M.S.)--Oregon Graduate Center, 1986.

Made available in DSpace on 2014-06-11T19:24:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-08-29Bitstream added on 2014-06-13T19:51:12Z : No. of bitstreams: 1 palu_jp_me_bauru.pdf: 727574 bytes, checksum: e4b549d779d54e15ec661bc6d8fae2ba (MD5) / Universidade Estadual Paulista (UNESP) / O presente trabalho tem o objetivo de estudar as relações no processo de montagem cinematográfica a partir da análise dos filmes Um homem com uma câmera (1929) de Dziga Vertov e Koyaanisqatsi (1983) de Godfrey Reggio. A escolha de tais filmes justifica-se, pois ambos procuram em sua montagem trabalhar a relação entre as imagens de uma maneira mais orgânica, relacional, não havendo tantos cortes que geram descontinuidade como na maioria dos filmes narrativos comerciais. A pesquisa também apresenta um panorama sobre o desenvolvimento e estabelecimento da linguagem cinematográfica desde seus pioneiros até o cinema captado com câmeras digitais, passando pelos grandes movimentos do século XX como, por exemplo, o Neorealismo italiano ou a Nouvelle Vague francesa e o quanto os filmes provenientes desses movimentos ajudaram no estabelecimento do cinema enquanto linguagem e expressão artística no século XX. A partir dos escritos teóricos dos grandes cineastas russos dos anos 1920-30 como Sergei Eisenstein, Vsevolod Pudovkin e Dziga Vertov, além dos estudiosos brasileiros Eduardo Leone e Maria Dora Mourão, apresenta-se um estudo sobre a importância da montagem no processo de construção de um filme. A análise do processo de montagem em Um homem com uma câmera e Koyaanisqatsi evidencia os pontos de contato entre ambos revelando uma lógica da linguagem fílmica, mesmo tendo sido realizados em um intervalo de mais de 50 anos e em duas culturas cinematográficas bastante distintas, além do que cada um tem de particular e o faz ser visto como produto de sua época / The present research has the objective of studying the relationship in the cinematographical editing process from the analyses of the films A man with a movie camera (1929) by Dziga Vertov and Koyaanisqatsi (1983) by Godfrey Reggio. The selection of such films is justified, because both try in its editing to work the relationship among the images in a more organical, relational way, there are not so many cuts which generate discontinuity as in most commercial narrative films. The research also presents a panorama about the development and establishment of the cinematographical language since its beginning until the cinema captured with the aid of digital cameras, passing through the greatest movements of the 20th century, for example, the Italian New-realism or the French Nouvelle Vague and how much the films originary of these movements helped in the establishment of the cinema as language and artistic expression in the 20th century. From the rhetorical writing of the greatest Russian moviemakers in the 1920´s and 1930´s like Sergei Eisenstein, Vsevolod Pudovkin and Dziga Vertov, besides the Brazilian researchers Eduardo Leone and Maria Dora Mourão, a study is presented about the importance of the editing in the process of construction of a film. The analysis of the editing process of A man with a movie camera and Koyaanisqatsi to evident the connecting points between them revealing a logic of the film language, even being produced more than 50 years apart each other and in two totally different cinematographic cultures, besides what each one presents as being of itself and what makes them be seen as products of their times