REGULATORY FUNCTIONS OF PROTEIN NUCLEIC ACID INTERACTION

蛋白质核酸相互作用的调节功能

• 批准号: 3093395
• 负责人: Wolfgang K Joklik
• 金额: $ 62.83万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-08-01 至 1987-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3093395
• 关键词: virus genetics; virus replication; virus virus interaction

Specific interactions between proteins and nucleic acids are the fundamental events that effect the expression, replication and packaging of genetic elements. This proposal describes studies on the molecular aspects of such interactions. Three systems are under investigation. A. We propose to examine the mechanisms involved in the generation of spontaneous rearrangements of the orthopoxvirus genome, focusing in particular on the factors that determine the extent, the sites and the frequency of such rearrangements. We will also study the control of viral transcription by identifying and characterizing strong late promoter elements in the cowpox virus genome. We will also attempt to identify the replication origins in cowpox virus DNA. B. We will continue our studies on mutants of the 37 Kd replication initiator protein of the plasmid pSC101; we will test these mutants for their ability to replicate. We will also express in E. coli and purify the pX protein of HTLV-1 and the protein encoded by the El open reading frame of BPV. We will use for this purpose an expression vector recently constructed by us, namely the collagen vector pJG200. We will also test these proteins for sequence specific DNA-binding ability. C. We will study the mechanisms involved in the rapid and complete inhibition of the processing and assembly of small nuclear ribonucleoproteins in cells infected with vesicular stomatitis virus. This effect is associated with the accumulation of precursor forms of U1 and U2 RNA. We will determine whether splicing of messenger RNAs in general is also inhibited. We will also continue our studies on the identification and function of proteins associated with ribosomal RNA and identify the genomic organization of the genes encoding these proteins. Finally, we will sequence the La protein gene; this is the protein with which VSV leader RNA interacts in infected cells. We will map the introns and exons of this gene, as well as identify the promoters and other upstream control sequences of its mRNA.

蛋白质和核酸之间的特定相互作用是 影响表达、复制和包装的基本事件 遗传元素。 该提案描述了分子方面的研究 的此类互动。 三个系统正在接受调查。 A. 我们建议研究涉及生成的机制 正痘病毒基因组的自发重排，集中于 特别是决定范围、地点和范围的因素 此类重排的频率。 我们还将研究病毒的控制 通过识别和表征强晚期启动子进行转录 牛痘病毒基因组中的元素。 我们还将尝试确定 复制起源于牛痘病毒DNA。 B. 我们将继续研究 37 Kd 复制突变体 质粒pSC101的起始蛋白；我们将测试这些突变体 他们的复制能力。 我们还将在大肠杆菌中表达并纯化 HTLV-1的pX蛋白和El开放阅读框编码的蛋白 BPV。 我们最近将为此目的使用表达载体 我们构建的胶原蛋白载体pJG200。 我们也会测试 这些蛋白质具有序列特异性 DNA 结合能力。 C. 我们将研究快速、完整的相关机制 抑制小核的加工和组装 水泡性口炎病毒感染细胞中的核糖核蛋白。 这 效应与 U1 和 U2 前体形式的积累有关 核糖核酸。 我们将确定信使 RNA 的剪接是否普遍存在 也受到抑制。 我们还将继续进行鉴定研究 和与核糖体 RNA 相关的蛋白质的功能，并确定 编码这些蛋白质的基因的基因组组织。 最后，我们 对La蛋白基因进行测序；这是 VSV 所用的蛋白质 前导 RNA 在受感染的细胞中相互作用。 我们将绘制内含子和外显子图 该基因，以及识别启动子和其他上游控制 其 mRNA 序列。