NOVEL CELL CYCLE REGULATION BY ADENOVIRUS E1B 55K

腺病毒 E1B 55K 对细胞周期的新型调控

• 批准号: 7909155
• 负责人: David Arnold Ornelles
• 金额: $ 13.56万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7909155
• 关键词: Accounting; Address; Adenovirus Infections; Adenoviruses; Attenuated; Cell Cycle; Cell Cycle Regulation; Cells; Clinical Trials; Complementary DNA; Defective Viruses; Employee Strikes; G1 Phase; Gene Expression; Genes; Goals; Growth; Human; Hybrid Cells; In Vitro; Infection; Malignant Neoplasms; Maps; Measures; Messenger RNA; Nature; Normal Cell; Oncolytic; Phase; Phase III Clinical Trials; Poly A; Protein Isoforms; Proteins; RNA Interference; RNA Splicing; RUNX1 gene; Recombinant DNA; Research; Research Personnel; Ribosomes; Rodent; Role; Scanning; Testing; Translating; Translations; Viral; Viral Genes; Viral Proteins; Virus; Virus Diseases; Work; base; human disease; killings; mutant; neoplastic cell; novel; overexpression; physical property; promoter; protein function; research study; response; tumor

DESCRIPTION (provided by applicant): Oncolytic ElB-55K-mutant adenoviruses have entered phase III clinical trials. However, the basis for their selective replication in tumor cells is poorly understood. Work of this lab has shown that ElB-55K-mutant viruses selectively replicate in and kill S phase cells better than G1 cells; this is likely to account for the selective replication in tumors. This research seeks to understand the tumor-selective nature of the E1B- 55K-mutant by understanding its S phase-selective and G1-restricted nature. Results from the previous period showed that late viral gene expression is suppressed in G1 cells at the level of translation, apparently in response to the E4orf1 or E4orf2 genes. The polysomal distribution of late viral mRNA in S phase and G1 cells infected with ElB-55K-mutant viruses suggests that late viral translation is restricted in G1 cells at the level of ribosome shunting. The E1B-55K protein of the wild-type virus promotes viral translation in association with the E4orl"6 protein and host cell factors. A candidate host cell factor has been mapped to the RUNX1 gene. Three specific aims are proposed in this application. Aim 1 seeks to determine how late viral translation is suppressed in G1-infected cells by comparing viral mRNA synthesis/ed in G1- and S phase-infected cells and by measuring the rates of key translational steps in synchronized and infected cells. This will test the hypothesis that ribosome shunting is decreased in the G1-infected cell. Aim 2 will elucidate the role of E4orfl and E4orf2 in attenuating late viral translation and restricting virus growth in G1. Viruses defective in either E4orlT or E4orf2 and the E1B-55K genes will be prepared and evaluated with respect to the cell cycle-restriction. Translation directed by ribosome scanning and ribosome shunting will he compared in cells infected with cell cycle-restricted and non-restricted mutant viruses to test the hypothesis that E4orf 1 attenuates translation at the level of ribosome shunting and that this is the basis for the G1-restriction. Aim 3 will identify the RUNX1 isoform important for E1B-55K function and determine if its activity during an infection is cell cycle-dependent. Candidate isoforms will be over expressed to determine if they compensate for the loss of E1B-55K function and RNA interference will be used to ablate RUNX1 expression to determine the importance of RUNX1 to adenovirus infection.

描述（由申请人提供）：溶瘤ELB-55K突变腺病毒已进入III期临床试验。但是，对它们在肿瘤细胞中的选择性复制的基础知之甚少。该实验室的工作表明，ELB-55K突变病毒选择性地复制并杀死S相细胞比G1细胞更好。这可能是肿瘤中选择性复制的原因。这项研究试图通过了解其相位选择性和G1限制性的性质来了解E1B-55K突变剂的肿瘤选择性。上一个时期的结果表明，在翻译水平的G1细胞中，晚期病毒基因表达显然是响应E4ORF1或E4ORF2基因的响应。晚期病毒mRNA在S期和感染ELB-55K突变病毒感染的G1细胞的多个分布表明，在核糖体分流水平的G1细胞中，晚期病毒翻译受到限制。野生型病毒的E1B-55K蛋白与E4orl“ 6蛋白和宿主细胞因子相关联）促进病毒翻译。候选宿主细胞因子已映射到Runx1基因。在此应用中提出了三个特定的目的。目的1寻求在G1中抑制的病毒型细胞，并在G1中抑制了G1的细胞，并在G1中抑制了G1的MRNA/MRNA SYNTONTON-MRNA SYNTHS SYNTHS SYNTONTH AINS SYNTHS SYNTONTH AINS SYNTONTH ins inn ins ins ins ins ins ins ins ins ins ins ins n.在同步和感染的细胞中测量关键的翻译步骤。对通过核糖体扫描和核糖体分流指导的细胞周期限制的评估将在感染细胞周期限制性和非限制性突变病毒的细胞中，以测试E4orf 1在核心切碎水平上减轻翻译的假设，这是G1限制的基础。 AIM 3将识别RUNX1同工型对于E1B-55K函数重要，并确定其在感染过程中的活性是否取决于细胞周期。候选同工型将过度表达，以确定它们是否补偿了E1B-55K功能的损失，并且RNA干扰将用于烧蚀Runx1表达式，以确定Runx1对腺病毒感染的重要性。

项目成果

Serotype-specific reorganization of the Mre11 complex by adenoviral E4orf3 proteins.

腺病毒 E4orf3 蛋白对 Mre11 复合物进行血清型特异性重组。

• DOI: 10.1128/jvi.79.11.6664-6673.2005
• 发表时间: 2005
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Stracker,TravisH;Lee,DarwinV;Carson,ChristianT;Araujo,FelipeD;Ornelles,DavidA;Weitzman,MatthewD
• 通讯作者: Weitzman,MatthewD

An activity associated with human chromosome 21 permits nuclear colocalization of the adenovirus E1B-55K and E4orf6 proteins and promotes viral late gene expression.

与人类 21 号染色体相关的活性允许腺病毒 E1B-55K 和 E4orf6 蛋白在核内共定位，并促进病毒晚期基因表达。

• DOI: 10.1128/jvi.77.14.8087-8098.2003
• 发表时间: 2003
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Chastain-Moore,AmyM;Roberts,Terry;Trott,DeborahA;Newbold,RobertF;Ornelles,DavidA
• 通讯作者: Ornelles,DavidA

Diverse roles for E4orf3 at late times of infection revealed in an E1B 55-kilodalton protein mutant background.

E1B 55 千道尔顿蛋白突变体背景揭示了 E4orf3 在感染后期的不同作用。

• DOI: 10.1128/jvi.78.18.9924-9935.2004
• 发表时间: 2004
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Shepard,RobinN;Ornelles,DavidA
• 通讯作者: Ornelles,DavidA