한빛사논문
- 조회1,949
Abstract
Daan C. Swarts1, 5, 6, Malwina Szczepaniak2, 5, Gang Sheng3, 5, Stanley D. Chandradoss2, Yifan Zhu1, Elizabeth M. Timmers1, Yong Zhang3, Hongtu Zhao3, Jizhong Lou3, Yanli Wang3, 4,*, Chirlmin Joo2,*, John van der Oost1, 7,*
1 Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, 6708 WE Wageningen, the Netherlands
2 Kavli Institute of NanoScience, Department of BioNanoScience, Delft University of Technology, 2628 CD Delft, the Netherlands
3 Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
4 University of Chinese Academy of Sciences, Beijing 100049, China
5 Co-first author
6 Present address: Department of Biochemistry, University of Zurich, 8006 Zurich, Switzerland
7 Lead Contact
*Corresponding authors
Summary
Several prokaryotic Argonaute proteins (pAgos) utilize small DNA guides to mediate host defense by targeting invading DNA complementary to the DNA guide. It is unknown how these DNA guides are being generated and loaded onto pAgo. Here, we demonstrate that guide-free Argonaute from Thermus thermophilus (TtAgo) can degrade double-stranded DNA (dsDNA), thereby generating small dsDNA fragments that subsequently are loaded onto TtAgo. Combining single-molecule fluorescence, molecular dynamic simulations, and structural studies, we show that TtAgo loads dsDNA molecules with a preference toward a deoxyguanosine on the passenger strand at the position opposite to the 5′ end of the guide strand. This explains why in vivo TtAgo is preferentially loaded with guides with a 5′ end deoxycytidine. Our data demonstrate that TtAgo can independently generate and selectively load functional DNA guides.
논문정보
- Research article
- 2017년 03월 (BRIC 등록일 2017-03-07)
- 국외 연구진
- 생명과학 > 구조생물 및 생물물리학
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