Direct observation and analysis of the dynamics of the photoresponsive transcription factor GAL4
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.
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.Attached Files
Accepted Version - Endo_et_al-2019-Angewandte_Chemie_International_Edition.pdf
Supplemental Material - downloadSupplement_doi=10.1002_2Fanie.201900610_file=anie201900610-s1-GAL4-VVD_Angew_Chem_revision_SI.pdf
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Supplemental Material - downloadSupplement_doi=10.1002_2Fanie.201900610_file=anie201900610-s4-Movie_S2.avi
Supplemental Material - downloadSupplement_doi=10.1002_2Fanie.201900610_file=anie201900610-s5-Movie_S3.avi
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Additional details
- Eprint ID
- 94239
- Resolver ID
- CaltechAUTHORS:20190328-093305414
- 18KK0139
- Japan Society for the Promotion of Science (JSPS)
- 16H06356
- Japan Society for the Promotion of Science (JSPS)
- Uehara Memorial Foundation
- Nakatani Foundation
- Kyoto University Foundation
- Honda-YES Foundation
- Created
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2019-03-28Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field