Functional and biophysical mapping of archaeal transcription complexes

古菌转录复合物的功能和生物物理图谱

• 批准号: BB/H019332/1
• 负责人: Finn Werner
• 金额: $ 54.18万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2010
• 资助国家: 英国
• 起止时间: 2010 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FH019332%2F1
• 关键词: Functional; biophysical; mapping; archaeal; transcription

The first step in gene expression is transcription of DNA into RNA and this is facilitated by RNA polymerases (RNAPs). RNAPs are large enzymes that are made of twelve subunits. We do currently not understand the function of all subunits but have a good working knowledge of many of their functions. RNAPs bind to their template (DNA), substrate (NTP) and product (RNA) and they interact with regulatory transcription factors. Much research on RNAPs is based on determining the structure of the enzyme. Large enzymes such as RNAP are dynamic and flexible, they often consist of a rigid frame and movable parts that can reorient themselves to accommodate interactions with e. g. DNA/RNA and regulatory factors. This flexibility is likely to be the underlying mechanical basis of RNAP function. However, even though many structural changes have been predicted it has not been possible to observe them directly. Our laboratory has developed methods to build RNAP from individual subunits. These can be labelled with fluorescent probes - molecular beacons that allow to measure distances and moreover changes in distances between them. We want to combine sophisticated fluorescence technology with our system to characterise for the first time the dynamics of RNAP.

基因表达的第一步是将DNA转录成RNA，这是由RNA聚合酶(RNAP)促进的。RNAP是由12个亚基组成的大型酶。我们目前不了解所有亚基的功能，但对它们的许多功能有很好的工作知识。RNAP与其模板(DNA)、底物(NTP)和产物(RNA)结合，并与调节转录因子相互作用。对RNAPs的许多研究都建立在确定酶的结构的基础上。像RNAP这样的大型酶是动态的和灵活的，它们通常由一个刚性的框架和可以重新定位的可移动部分组成，以适应与DNA/RNA和调节因子的相互作用。这种灵活性很可能是RNAP功能的基本机械基础。然而，即使预测到了许多结构性变化，也不可能直接观察到它们。我们的实验室已经开发出从单个亚基构建RNAP的方法。这些可以用荧光探针标记--分子信标允许测量距离，而且还可以测量它们之间的距离变化。我们希望将复杂的荧光技术与我们的系统相结合，首次表征RNAP的动力学。

项目成果

The initiation factor TFE and the elongation factor Spt4/5 compete for the RNAP clamp during transcription initiation and elongation.

• DOI: 10.1016/j.molcel.2011.05.030
• 发表时间: 2011-07-22
• 期刊: MOLECULAR CELL
• 影响因子: 16
• 作者: Grohmann, Dina;Nagy, Julia;Chakraborty, Anirban;Klose, Daniel;Fielden, Daniel;Ebright, Richard H.;Michaelis, Jens;Werner, Finn
• 通讯作者: Werner, Finn

Evolutionary history of the TBP-domain superfamily.

• DOI: 10.1093/nar/gkt045
• 发表时间: 2013-03-01
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Brindefalk B;Dessailly BH;Yeats C;Orengo C;Werner F;Poole AM
• 通讯作者: Poole AM

Lytic water dynamics reveal evolutionarily conserved mechanisms of ATP hydrolysis by TIP49 AAA+ ATPases.

• DOI: 10.1016/j.str.2014.02.002
• 发表时间: 2014-04-08
• 期刊: Structure (London, England : 1993)
• 作者: Afanasyeva A;Hirtreiter A;Schreiber A;Grohmann D;Pobegalov G;McKay AR;Tsaneva I;Petukhov M;Käs E;Grigoriev M;Werner F
• 通讯作者: Werner F