GENETICS OF PRIMATE D TYPE RETROVIRUSES

灵长类 D 型逆转录病毒的遗传学

• 批准号: 6322351
• 负责人: Eric Hunter
• 金额: $ 6.11万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2003-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6322351
• 关键词: Mason Pfizer monkey virus; X ray crystallography; active sites; capsid; crystallization; endopeptidases; enzyme mechanism; enzyme structure; fatty acylation; glycoprotein structure; intracellular transport; molecular cloning; mutant; myristates; posttranslational modifications; protein structure function; protein transport; proteolysis; virus assembly; virus genetics; virus protein; virus replication

The proposed studies aim to understand the molecular mechanisms governing assembly and maturation of retroviral capsids, specifically the role of the matrix (MA) protein and protease (PR). Previous studies by the PI with his foreign collaborators, using Mason-Pfizer Monkey virus (M-PMV) as a model system, have shown the assembly of retroviral capsids from the gag precursors in vitro after over expression of this protein in bacteria and a new-form of the proteinase. The proposed studies will extend these observations to identify the domains of gag required for capsid targeting and assembly (specific aim 1) and to study the proteolytic process of the gag polyproteins (specific aim 2). The subgoals for these specific aims are to: la. Clone and express the M- PMV membrane targeting defective MA mutants; 1b. Determine the structure of the mutant MA protein and compare its structure with the wild type protein to elucidate the importance of specific domains in structural changes that affect intracellular targeting, assembly, and transport; 1c. Study the effect of N-terminal myristylation on MA protein structure; 1d. Express and purify p12 and p16 for structural analyses; 1e. Study the role of the MA domain in assembly of immature capsids using chimeric gag genes from C and D-type retroviruses; 1f. Determine the role of the gag-encoded N-terminal domains for efficient assembly of immature capsids; 2a. Study the role of the truncated proteinase species in the viral replication cycle; 2b. Study the substrate specificity of the proteinase using the gag polyprotein; 2c. Screen for inhibitors of the proteinase using a combinatorial peptide library; 2d. Crystallize and determine the structure of the different forms of the proteinase.

拟议的研究旨在了解分子机制 逆转录病毒包装的管理和成熟，特别是 基质（MA）蛋白质和蛋白酶（PR）的作用。 先前的研究 PI与他的外国合作者使用Mason-Pfizer Monkey 病毒（M-PMV）作为模型系统，已显示逆转录病毒的组装 过度表达后，在体外插入cag前体的衣壳 细菌中的蛋白质和一种新的蛋白酶。 提议 研究将扩展这些观察结果以识别GAG的领域 capsID靶向和组装所需（特定目标1）和研究 GAG多蛋白的蛋白水解过程（特定目标2）。这 这些特定目标的子目标是：la。 克隆并表达M- PMV膜靶向有缺陷的MA突变体； 1B。确定结构 突变MA蛋白的结构与野生型进行比较 蛋白质以阐明特定域在结构中的重要性 影响细胞内靶向，组装和运输的变化； 1C。研究N端肉豆蔻酰化对MA蛋白的影响 结构; 1d表达和净化p12和p16进行结构分析； 1e。研究MA结构域在未成熟衣壳组装中的作用 使用来自C和D型逆转录病毒的嵌合GAG基因； 1f。决定 GAG编码的N末端域在有效组装中的作用 未成熟的衣壳； 2a。研究截短的蛋白酶的作用 病毒复制周期中的物种； 2b。研究底物 使用GAG多蛋白的蛋白酶的特异性； 2C。屏幕 使用联合肽库的蛋白酶抑制剂； 2d。 结晶并确定不同形式的结构 蛋白酶。

项目成果

Phagemid pSIT permits efficient in vitro mutagenesis and tightly controlled expression in E. coli.

噬菌粒 pSIT 允许在大肠杆菌中进行有效的体外诱变和严格控制的表达。

• 发表时间: 1994
• 期刊: BioTechniques
• 影响因子: 2.7
• 作者: Andreansky,M;Hunter,E
• 通讯作者: Hunter,E

An investigation of sociomedical risk factors associated with vaginal fistula in northern Nigeria.

尼日利亚北部与阴道瘘相关的社会医学危险因素的调查。

• DOI: 10.1300/j013v28n03_07
• 发表时间: 1999
• 期刊: Women & health
• 影响因子: 1.6
• 作者: OjanugaOnolemhemhen,D;Ekwempu,CC
• 通讯作者: Ekwempu,CC

A pmr study of the effects of pH and anion and metal ion binding of the histidyl residues of ovotransferrin.

PMR 研究 pH 值以及阴离子和金属离子对卵转铁蛋白组氨酰残基结合的影响。

• DOI: 10.1016/s0162-0134(00)80131-2
• 发表时间: 1981
• 期刊: Journal of inorganic biochemistry
• 影响因子: 3.9
• 作者: Alsaadi,BM;Williams,RJ;Woodworth,RC
• 通讯作者: Woodworth,RC

Backbone resonance assignment of protease from Mason-Pfizer monkey virus.

梅森-辉瑞猴病毒蛋白酶的主链共振分配。

• DOI: 10.1023/a:1011202229203
• 发表时间: 2001
• 期刊: Journal of biomolecular NMR
• 影响因子: 2.7
• 作者: Veverka,V;Bauerová,H;Zábranský,A;Pichová,I;Hrabal,R
• 通讯作者: Hrabal,R

Three active forms of aspartic proteinase from Mason-Pfizer monkey virus.

梅森-辉瑞猴病毒天冬氨酸蛋白酶的三种活性形式。

• DOI: 10.1006/viro.1998.9173
• 发表时间: 1998
• 期刊: Virology.
• 作者: Zabransky,A;Andreansky,M;Hruskova-Heidingsfeldova,O;Havlicek,V;Hunter,E;Ruml,T;Pichova,I
• 通讯作者: Pichova,I