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Auto-inhibition and activation of a short Argonaute-associated TIR-APAZ defense system

Nat Chem Biol. 2024-04; 
Lijie Guo, Pingping Huang, Zhaoxing Li, Young-Cheul Shin, Purui Yan, Meiling Lu, Meirong Chen, Yibei Xiao
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Molecular Biology Reagents … thermophila TIR-APAZ and short Ago were chemically synthesized (GenScript) and cloned into the vector pCOLADuet-1 using homologous recombination, with the 6×His tag attached … Get A Quote
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摘要

Short prokaryotic Ago accounts for most prokaryotic Argonaute proteins (pAgos) and is involved in defending bacteria against invading nucleic acids. Short pAgo associated with TIR-APAZ (SPARTA) has been shown to oligomerize and deplete NAD upon guide-mediated target DNA recognition. However, the molecular basis of SPARTA inhibition and activation remains unknown. In this study, we determined the cryogenic electron microscopy structures of Crenotalea thermophila SPARTA in its inhibited, transient and activated states. The SPARTA monomer is auto-inhibited by its acidic tail, which occupies the guide-target binding channel. Guide-mediated target binding expels this acidic tail and triggers substantial conformation... More

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