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Direct observation and analysis of the dynamics of the photoresponsive transcription factor GAL4

Raghavan, Guruprasad and Hidaka, Kumi and Sugiyama, Hiroshi and Endo, Masayuki (2019) Direct observation and analysis of the dynamics of the photoresponsive transcription factor GAL4. Angewandte Chemie International Edition, 58 (23). pp. 7626-7630. ISSN 1433-7851. https://resolver.caltech.edu/CaltechAUTHORS:20190328-093305414

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Abstract

Herein, the direct visualization of the dynamic interaction between a photoresponsive transcription factor fusion, GAL4–VVD, and DNA using high‐speed atomic force microscopy (HS‐AFM) is reported. A series of different GAL4–VVD movements, such as binding, sliding, stalling, and dissociation, was observed. Inter‐strand jumping on two double‐stranded (ds) DNAs was also observed. Detailed analysis using a long substrate DNA strand containing five GAL4‐binding sites revealed that GAL4–VVD randomly moved on the dsDNA using sliding and hopping to rapidly find specific binding sites, and then stalled to the specific sites to form a stable complex formation. These results suggest the existence of different conformations of the protein to enable sliding and stalling. This single‐molecule imaging system with nanoscale resolution provides an insight into the searching mechanism used by DNA‐binding proteins.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1002/anie.201900610DOIArticle
Additional Information:© 2019 WILEY‐VCH. Manuscript received: 16 January 2019. Manuscript revised: 11 March 2019. Manuscript accepted: 14 March 2019. Accepted manuscript online: 25 March 2019. This work was supported by a Grant‐in‐Aid for Scientific Research JSPS KAKENHI Fund for the Promotion of Joint International Research (Fostering Joint International Research (B), Grant Numbers: 18KK0139 and 16H06356) to M.E. Financial support from the Uehara Memorial Foundation, the Nakatani Foundation, and the Kyoto University Foundation to M.E. are acknowledged. G.P.R. also acknowledges the Honda‐YES Foundation for supporting his stay at Kyoto University. The authors declare no conflict of interest.
Funders:
Funding AgencyGrant Number
Japan Society for the Promotion of Science (JSPS)18KK0139
Japan Society for the Promotion of Science (JSPS)16H06356
Uehara Memorial FoundationUNSPECIFIED
Nakatani FoundationUNSPECIFIED
Kyoto University FoundationUNSPECIFIED
Honda-YES FoundationUNSPECIFIED
Subject Keywords:DNA origami • transcription factor • photo-control • single-molecule observation• high-speed AFM
Issue or Number:23
Record Number:CaltechAUTHORS:20190328-093305414
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20190328-093305414
Official Citation:G. Raghavan, K. Hidaka, H. Sugiyama, M. Endo, Angew. Chem. Int. Ed. 2019, 58, 7626; doi: 10.1002/anie.201900610
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:94239
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:28 Mar 2019 18:08
Last Modified:03 Oct 2019 21:02

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