DNA CONFORMATION AND PROTEIN-DNA INTERACTION

DNA 构象和蛋白质-DNA 相互作用

• 批准号: 3283420
• 负责人: ROGER M WARTELL
• 金额: $ 8.28万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-09-01 至 1988-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3283420
• 关键词: DNA; DNA directed RNA polymerase; Raman spectrometry; bacteria; bacterial DNA; circular dichroism; conformation; cyclic AMP; endonuclease; genetic manipulation; genetic transcription; lac operon; microorganism genetics; nucleic acid reconstitution; nucleobase; spectrometry

The goal of this proposal is to contribute to the understanding of transcription initiation from the lactose (lac) operon of Escherichia coli. The first objective of the proposal is to obtain large quantities of two DNA fragments which contain the binding sites for RNA polymerase and the catabolite activator protein - cyclic adenosine monophosphate complex (CAP.cAMP). This will be done by growing up cells containing plasmids with the DNA fragments inserted into a single restriction endonuclease site, and cutting out and isolating the DNA fragments. These DNAs will be combined and altered to generate a modified promoter DNA with nucleotide analogues at specific locations near the transcription startpoint. Biophysical and Biochemical studies will be carried out on the DNAs. The influence of nucleotide replacements on RNA polymerase binding and transcription will be examined. Raman spectroscopy will be employed to examine the conformation of the unmodified lac DNA promoter and to determine the effect of CAP.cAMP binding on the vibrational properties of base pairs around the startpoint region.

这个建议的目的是有助于理解转录 由大肠杆菌的乳糖操纵子起始。 第一 该提案的目的是获得大量的两个DNA片段， 其含有RNA聚合酶和分解代谢物激活剂的结合位点 蛋白质-环磷酸腺苷复合物（CAP.cAMP）。 为此将 通过培养含有质粒的细胞，质粒中的DNA片段插入到 单个限制性内切酶位点，切下并分离DNA 片段 这些DNA将被组合和改变，以产生一个修改后的 启动子DNA与核苷酸类似物在特定位置附近的 转录起始点。 将对DNA进行生物物理和生物化学研究。 的 核苷酸替换对RNA聚合酶结合和转录影响 将被审查。 将采用拉曼光谱来检查 未修饰的lac DNA启动子的构象，并确定 CAP.cAMP结合对周围碱基对振动特性的影响 起始点区域。

项目成果

The catabolite activator protein stabilizes its binding site in the E. coli lactose promoter.

分解代谢物激活蛋白稳定其在大肠杆菌乳糖启动子中的结合位点。

• DOI: 10.1093/nar/13.20.7483
• 发表时间: 1985
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: DeGrazia,H;Abhiraman,S;Wartell,RM
• 通讯作者: Wartell,RM

Intensity changes of base and backbone Raman modes in the B to Z transition of poly(dG-dm5C).

聚 (dG-dm5C) B 到 Z 转变中基础和主链拉曼模式的强度变化。

• DOI: 10.1080/07391102.1986.10508494
• 发表时间: 1986
• 期刊: Journal of biomolecular structure & dynamics
• 影响因子: 4.4
• 作者: Brown,DM;Wartell,RM
• 通讯作者: Wartell,RM

DNA bending induced by the catabolite activator protein allows ring formation of a 144 bp DNA.

分解代谢物激活蛋白诱导的 DNA 弯曲允许形成 144 bp DNA 环。

• DOI: 10.1080/07391102.1987.10506370
• 发表时间: 1987
• 期刊: Journal of biomolecular structure & dynamics
• 影响因子: 4.4
• 作者: Dripps,D;Wartell,RM
• 通讯作者: Wartell,RM

Characteristics and variations of B-type DNA conformations in solution: a quantitative analysis of Raman band intensities of eight DNAs.

溶液中 B 型 DNA 构象的特征和变化：八种 DNA 拉曼带强度的定量分析。

• DOI: 10.1021/bi00357a056
• 发表时间: 1986
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Wartell,RM;Harrell,JT
• 通讯作者: Harrell,JT

Solution conformations of DNAs containing binding sites of the catabolite gene activator protein and lac repressor protein: characterization by Raman spectroscopy.

含有分解代谢物基因激活蛋白和 lac 阻遏蛋白结合位点的 DNA 的溶液构象：拉曼光谱表征。

• DOI: 10.1021/bi00417a025
• 发表时间: 1988
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: DeGrazia,H;Brown,D;Cheung,S;Wartell,RM
• 通讯作者: Wartell,RM