Conformational changes and catalytic inefficiency associated with Mot1-mediated TBP–DNA dissociation
| Autor: | David T. Auble, Ramya Viswanathan, Michael Meisterernst, Samson Glaser, Sushi Madhira, Lena Voith von Voithenberg, Peter Schluesche, Gregor Heiss, Don C. Lamb, Evelyn Ploetz |
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| Rok vydání: | 2019 |
| Předmět: |
Models
Molecular Conformational change Saccharomyces cerevisiae Proteins Protein Conformation genetic processes Saccharomyces cerevisiae macromolecular substances Biology environment and public health Catalysis 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Protein structure ATP hydrolysis Transcription (biology) Gene Expression Regulation Fungal Escherichia coli Genetics DNA Fungal Promoter Regions Genetic 030304 developmental biology Adenosine Triphosphatases TATA-Binding Protein Associated Factors 0303 health sciences Gene regulation Chromatin and Epigenetics Promoter TATA-Box Binding Protein enzymes and coenzymes (carbohydrates) Förster resonance energy transfer chemistry health occupations Biophysics Nucleic Acid Conformation Adenosine triphosphate 030217 neurology & neurosurgery DNA Protein Binding |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gky1322 |
| Popis: | The TATA-box Binding Protein (TBP) plays a central role in regulating gene expression and is the first step in the process of pre-initiation complex (PIC) formation on promoter DNA. The lifetime of TBP at the promoter site is controlled by several cofactors including the Modifier of transcription 1 (Mot1), an essential TBP-associated ATPase. Based on ensemble measurements, Mot1 can use adenosine triphosphate (ATP) hydrolysis to displace TBP from DNA and various models for how this activity is coupled to transcriptional regulation have been proposed. However, the underlying molecular mechanism of Mot1 action is not well understood. In this work, the interaction of Mot1 with the DNA/TBP complex was investigated by single-pair Förster resonance energy transfer (spFRET). Upon Mot1 binding to the DNA/TBP complex, a transition in the DNA/TBP conformation was observed. Hydrolysis of ATP by Mot1 led to a conformational change but was not sufficient to efficiently disrupt the complex. SpFRET measurements of dual-labeled DNA suggest that Mot1’s ATPase activity primes incorrectly oriented TBP for dissociation from DNA and additional Mot1 in solution is necessary for TBP unbinding. These findings provide a framework for understanding how the efficiency of Mot1’s catalytic activity is tuned to establish a dynamic pool of TBP without interfering with stable and functional TBP-containing complexes. |
| Databáze: | OpenAIRE |
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