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Asterix/Gtsf1 links tRNAs and piRNA silencing of retrotransposons

Cell Rep. 2021-03; 
Jonathan J Ipsaro, Paul A O'Brien, Shibani Bhattacharya, Arthur G Palmer, Leemor Joshua-Tor
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Insect Expression  coli or insect cell culture systems. In addition, natural sequences of the D. melanogaster and M. musculus proteins were used for transfection in eCLIP experiments. The sequence of each construct was verified by GenScript Get A Quote

摘要

The Piwi-interacting RNA (piRNA) pathway safeguards genomic integrity by silencing transposable elements (transposons) in the germline. While Piwi is the central piRNA factor, others including Asterix/Gtsf1 have also been demonstrated to be critical for effective silencing. Here, using enhanced crosslinking and immunoprecipitation (eCLIP) with a custom informatic pipeline, we show that Asterix/Gtsf1 specifically binds tRNAs in cellular contexts. We determined the structure of mouse Gtsf1 by NMR spectroscopy and identified the RNA-binding interface on the protein's first zinc finger, which was corroborated by biochemical analysis as well as cryo-EM structures of Gtsf1 in complex with co-purifying tRNA. Consisten... More

关键词

Asterix, Gtsf1, LTR, NMR, RNA, cryoEM, eCLIP, piRNA, retrotransposon, tRNA