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TitleStructural RNA components supervise the sequential DNA cleavage in R2 retrotransposon.
Journal, issue, pagesCell, Vol. 186, Issue 13, Page 2865-2879.e20, Year 2023
Publish dateJun 22, 2023
AuthorsPujuan Deng / Shun-Qing Tan / Qi-Yu Yang / Liangzheng Fu / Yachao Wu / Han-Zhou Zhu / Lei Sun / Zhangbin Bao / Yi Lin / Qiangfeng Cliff Zhang / Haoyi Wang / Jia Wang / Jun-Jie Gogo Liu /
PubMed AbstractRetroelements are the widespread jumping elements considered as major drivers for genome evolution, which can also be repurposed as gene-editing tools. Here, we determine the cryo-EM structures of ...Retroelements are the widespread jumping elements considered as major drivers for genome evolution, which can also be repurposed as gene-editing tools. Here, we determine the cryo-EM structures of eukaryotic R2 retrotransposon with ribosomal DNA target and regulatory RNAs. Combined with biochemical and sequencing analysis, we reveal two essential DNA regions, Drr and Dcr, required for recognition and cleavage. The association of 3' regulatory RNA with R2 protein accelerates the first-strand cleavage, blocks the second-strand cleavage, and initiates the reverse transcription starting from the 3'-tail. Removing 3' regulatory RNA by reverse transcription allows the association of 5' regulatory RNA and initiates the second-strand cleavage. Taken together, our work explains the DNA recognition and RNA supervised sequential retrotransposition mechanisms by R2 machinery, providing insights into the retrotransposon and application reprogramming.
External linksCell / PubMed:37301196
MethodsEM (single particle)
Resolution3.04 - 3.74 Å
Structure data

EMDB-35347, PDB-8ibw:
Structure of R2 with 3'UTR and DNA in binding state
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-35348, PDB-8ibx:
Structure of R2 with 3'UTR and DNA in unwinding state
Method: EM (single particle) / Resolution: 3.74 Å

EMDB-35349, PDB-8iby:
Structure of R2 with 5'ORF
Method: EM (single particle) / Resolution: 3.47 Å

EMDB-35350, PDB-8ibz:
Structure of R2 with 5'ORF and 3'UTR
Method: EM (single particle) / Resolution: 3.04 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • bombyx mori (domestic silkworm)
KeywordsRNA BINDING PROTEIN/RNA/DNA / R2 complex / RNA BINDING PROTEIN-RNA-DNA complex / RNA BINDING PROTEIN/RNA / RNA BINDING PROTEIN-RNA complex

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