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TitleAuto-inhibition and activation of a short Argonaute-associated TIR-APAZ defense system.
Journal, issue, pagesNat Chem Biol, Vol. 20, Issue 4, Page 512-520, Year 2024
Publish dateNov 6, 2023
AuthorsLijie Guo / Pingping Huang / Zhaoxing Li / Young-Cheul Shin / Purui Yan / Meiling Lu / Meirong Chen / Yibei Xiao /
PubMed AbstractShort 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 ...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 conformational changes to expose the Ago-Ago dimerization interface. As a result, SPARTA assembles into an active tetramer, where the four TIR domains are rearranged and packed to form NADase active sites. Together with biochemical evidence, our results provide a panoramic vision explaining SPARTA auto-inhibition and activation and expand understanding of pAgo-mediated bacterial defense systems.
External linksNat Chem Biol / PubMed:37932527
MethodsEM (single particle)
Resolution3.3 - 4.2 Å
Structure data

36059

8j84

EMDB-36059, PDB-8j84:
Short ago complexed with TIR-APAZ
Method: EM (single particle) / Resolution: 3.3 Å

36070

8j8h

EMDB-36070, PDB-8j8h:
SPARTA monomer bound with guide-target, state 2
Method: EM (single particle) / Resolution: 3.4 Å

36095

8j9g

EMDB-36095, PDB-8j9g:
CrtSPARTA hetero-dimer bound with guide-target, state 1
Method: EM (single particle) / Resolution: 3.5 Å

36114

8j9p

EMDB-36114, PDB-8j9p:
SPARTA dimer bound with guide-target
Method: EM (single particle) / Resolution: 3.4 Å

36138

8jay

EMDB-36138, PDB-8jay:
CrtSPARTA Octamer bound with guide-target
Method: EM (single particle) / Resolution: 4.2 Å

Chemicals

ChemComp-MG:
Unknown entry

Source
  • thermoflavifilum thermophilum (bacteria)
KeywordsDNA BINDING PROTEIN / SPARTA / Ago / Tir / DNA BINDING PROTEIN/DNA/RNA / DNA BINDING PROTEIN-DNA-RNA complex / DNA BINDING PROTEIN/RNA/DNA / DNA BINDING PROTEIN-RNA-DNA complex

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