[{"key":"dc.contributor.author","value":"Pathakamuri, Joseph Aja","language":null},{"key":"dc.date.accessioned","value":"2009-11-27T00:00:00","language":null},{"key":"dc.date.available","value":"2009-11-27T00:00:00","language":null},{"key":"dc.date.issued","value":"2004","language":null},{"key":"dc.identifier.uri","value":"http:\/\/hdl.handle.net\/2429\/15828","language":null},{"key":"dc.description.abstract","value":"Baculovirus Orgyia pseudotsugata multiple nucleopolyhedrovirus (OpMNPV) transient\r\nreplication assays have previously identified six genes as essential iel, lef-1, lef-2, lef-3, DNA\r\npol, helicase; three genes as stimulatory ie2, p34, iap-1; and two types of replication origins hrs\r\nand non-hrs. One of these essential proteins IE1, has an acidic activation domain (AAD) at the\r\nN-terminus that is required for transcriptional transactivation but its role in viral DNA\r\nreplication was unknown. In this study we have determined that the AAD is also essential for\r\nDNA replication. Unlike transcriptional activation, the AAD cannot be replaced with the\r\nAcMNPV IE1 and herpesvirus VP 16 AADs for replication. Deletion analysis of the OpMNPV\r\nIE 1-AAD revealed the presence of separate sub-domains for replication and transcriptional\r\nactivation. By substituting OpMNPV lef-1, -2, -3 and helicase proteins (part of the putative\r\nreplisome) with corresponding AcMNPV factors, the inactive OpMNPV IE1 chimeric protein\r\nIE 1-Ac AD, which contains the AcMNPV IE1 AAD, was functional for replication activation.\r\nThis suggests that AADs interact with viral replication factors in order to maintain viral\r\nspecificity in replication. The IE 1-Ac AD specificity for the AcMNPV replisome was found to\r\nbe similar to the native AcMNPV IE1, thus demonstrating that by changing the AAD the\r\nspecificity of the protein changes. Further studies showed that the role of stimulatory factor\r\nOpMNPV IE2 in replication was to maintain the specificity between AAD and the replisome.\r\nAbsence of IE2 allowed replication from non-specific AAD and replisome but presence of IE2\r\nactive replication was observed only between specific AAD and replisome.\r\nAcMNPV non-hr origin analysis indicated that it was unable to allow for replication with\r\nOpMNPV IE1 and OpMNPV replication proteins; however substituting with AcMNPV LEF-3,\r\nPOL and HELICASE, resulted in replication. This result reveals the presence of specific\r\ninteractions between origins and replication factors.\r\n\r\nOur studies on OpMNPV IEO, the only spliced gene of IE1, showed that IEO is functionally\r\nactive for replication and can replace IE1. We also performed initial functional analysis of\r\nOpMNPV EXONO. To date there is no information on OpMNPV EXONO function other than\r\nthe fact that it contributes 23aa to the N-terminus of IEO. Our study shows EXONO has a\r\nconserved novel ring finger and is expressed as a late gene. However, our preliminary research\r\ncould not identify any function attributable to EXONO.","language":"en"},{"key":"dc.format.extent","value":"14584839 bytes","language":null},{"key":"dc.format.mimetype","value":"application\/pdf","language":null},{"key":"dc.language.iso","value":"eng","language":"en"},{"key":"dc.publisher","value":"University of British Columbia","language":null},{"key":"dc.rights","value":"For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https:\/\/open.library.ubc.ca\/terms_of_use.","language":null},{"key":"dc.title","value":"Analysis of the orgyia pseudotsugata multiple nucleopolyhedrovirus IE1 acidic activation domain role in viral DNA replication","language":"en"},{"key":"dc.type","value":"Text","language":null},{"key":"dc.degree.name","value":"Doctor of Philosophy - PhD","language":"en"},{"key":"dc.degree.discipline","value":"Plant Science","language":"en"},{"key":"dc.degree.grantor","value":"University of British Columbia","language":null},{"key":"dc.date.graduation","value":"2004-05","language":"en"},{"key":"dc.type.text","value":"Thesis\/Dissertation","language":"en"},{"key":"dc.description.affiliation","value":"Land and Food Systems, Faculty of","language":null},{"key":"dc.degree.campus","value":"UBCV","language":"en"},{"key":"dc.description.scholarlevel","value":"Graduate","language":"en"}]