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-Structure paper
タイトル | Target ssDNA activates the NADase activity of prokaryotic SPARTA immune system. |
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ジャーナル・号・ページ | Nat Chem Biol, Vol. 20, Issue 4, Page 503-511, Year 2024 |
掲載日 | 2023年11月6日 |
著者 | Jun-Tao Zhang / Xin-Yang Wei / Ning Cui / Ruilin Tian / Ning Jia / |
PubMed 要旨 | Argonaute proteins (Agos), which use small RNAs or DNAs as guides to recognize complementary nucleic acid targets, mediate RNA silencing in eukaryotes. In prokaryotes, Agos are involved in immunity: ...Argonaute proteins (Agos), which use small RNAs or DNAs as guides to recognize complementary nucleic acid targets, mediate RNA silencing in eukaryotes. In prokaryotes, Agos are involved in immunity: the short prokaryotic Ago/TIR-APAZ (SPARTA) immune system triggers cell death by degrading NAD in response to invading plasmids, but its molecular mechanisms remain unknown. Here we used cryo-electron microscopy to determine the structures of inactive monomeric and active tetrameric Crenotalea thermophila SPARTA complexes, revealing mechanisms underlying SPARTA assembly, RNA-guided recognition of target single-stranded DNA (ssDNA) and subsequent SPARTA tetramerization, as well as tetramerization-dependent NADase activation. The small RNA guides Ago to recognize its ssDNA target, inducing SPARTA tetramerization via both Ago- and TIR-mediated interactions and resulting in a two-stranded, parallel, head-to-tail TIR rearrangement primed for NAD hydrolysis. Our findings thus identify the molecular basis for target ssDNA-mediated SPARTA activation, which will facilitate the development of SPARTA-based biotechnological tools. |
リンク | Nat Chem Biol / PubMed:37932528 |
手法 | EM (単粒子) |
解像度 | 2.54 - 3.11 Å |
構造データ | EMDB-35419, PDB-8ifk: EMDB-35420: Cryo-EM structure of tetrameric SPARTA gRNA-ssDNA target complex in state1 EMDB-35421, PDB-8ifm: EMDB-36843, PDB-8k34: |
化合物 | ChemComp-MG: |
由来 |
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キーワード | RNA BINDING PROTEIN/RNA/DNA / RNA BINDING PROTEIN-RNA-DNA COMPLEX / STRUCTURAL PROTEIN/RNA / STRUCTURAL PROTEIN-RNA COMPLEX |