期刊
ACS NANO
卷 4, 期 10, 页码 5969-5977出版社
AMER CHEMICAL SOC
DOI: 10.1021/nn101662a
关键词
atomic force microscopy (AFM); functionalized 2D origami; human topoisomerase I; secondary binding site; supercoil recognition
类别
资金
- Danish National Research Foundation
- Danish Ministry for Science, Technology, and Innovation through the iNANO Center
- Danish Research Councils
- Danish Cancer Society
- Novo Nordisk Foundation
- Carlsberg Foundation
- Augustinus Foundation
- Civilingenior Frode V. Nyegaard og hustru's Foundation
- Direktor Einar Hansen og hustru fru Vera Hansen's Foundation
- Fabrikant Einar Willumsen's Mindelegat
- Harboe Foundation
- Karen Elise Jensen's Foundation
- Kobmand Sven Hansen og hustru Ina Hansen's Fondation
- Aage og Johanne Louis-Hansens Foundation
- Horslev Foundation
- Foundation til Laegvidenskabens fremme
The biologically and clinically important nuclear enzyme human topoisomerase I relaxes both positively and negatively supercoiled DNA and binds consequently DNA with supercoils of positive or negative sign with a strong preference over relaxed DNA. One scheme to explain this preference relies on the existence of a secondary DNA binding site in the enzyme facilitating binding to DNA nodes characteristic for plectonemic DNA. Here we demonstrate the ability of human topoisomerase I to induce formation of DNA synapses at protein containing nodes or filaments using atomic force microscopy imaging. By means of a two-dimensional (2D) DNA origami platform, we monitor the interactions between a single human topoisomerase I covalently bound to one DNA fragment and a second DNA fragment protruding from the DNA origami. This novel single molecule origami-based detection scheme provides direct evidence for the existence of a secondary DNA interaction site in human topoisomerase I and lends further credence to the theory of two distinct DNA interaction sites in human topoisomerase I, possibly facilitating binding to DNA nodes characteristic for plectonemic supercoils.
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