Autonomous Generation and Loading of DNA Guides by Bacterial Argonaute

• Published in: Molecular cell Vol. 65; no. 6; pp. 985 - 998.e6
• Main Authors: Swarts, Daan C., Szczepaniak, Malwina, Sheng, Gang, Chandradoss, Stanley D., Zhu, Yifan, Timmers, Elizabeth M., Zhang, Yong, Zhao, Hongtu, Lou, Jizhong, Wang, Yanli, Joo, Chirlmin, van der Oost, John
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.03.2017
• Subjects: Ago; Argonaute Proteins - chemistry; Argonaute Proteins - genetics; Argonaute Proteins - metabolism; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Binding Sites; Deoxycytidine - metabolism; Deoxyguanosine - metabolism; DNA chopping; DNA, Antisense - chemistry; DNA, Antisense - genetics; DNA, Antisense - metabolism; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; DNA, Bacterial - metabolism; Fluorescence Resonance Energy Transfer; Guide generation; Guide loading; Molecular Dynamics Simulation; Nucleic Acid Conformation; PAgo; Prokaryotic argonaute; Protein Binding; Protein Conformation; RNA interference; SiDNA; Single Molecule Imaging; Small interfering DNA; Structure-Activity Relationship; Thermus thermophilus - enzymology; Thermus thermophilus - genetics; TtAgo; RNA interference; guide loading; small interfering DNA; siDNA; DNA chopping; ago; pAgo; prokaryotic argonaute; TtAgo; guide generation

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 thermophi...