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Dissection of purified LINE-1 reveals distinct nuclear and cytoplasmic intermediates

By K.R. Molloy, M.S. Taylor, Ilya Altukhov, P. Mita, H. Jiang, E.M. Adney, A. Wudzinska, D. Ischenko, K.H. Burns, David Fenyƶ, B.T. Chait, D. Alexeev, M.P. Rout, Jef D Boeke, John A LaCava

Posted 30 Jun 2017
bioRxiv DOI: 10.1101/157818 (published DOI: 10.7554/eLife.30094)

Long Interspersed Nuclear Element-1 (LINE-1, L1) is a mobile genetic element active in human genomes. L1-encoded ORF1 and ORF2 proteins bind L1 RNAs, forming ribonucleoproteins (RNPs). These RNPs interact with diverse host proteins, some repressive and others required for the L1 lifecycle. Using differential affinity purifications and quantitative mass spectrometry, we have characterized the proteins associated with distinctive L1 macromolecular complexes. Our findings support the presence of multiple L1-derived retrotransposition intermediates in vivo. Among them, we describe a cytoplasmic intermediate that we hypothesize to be the canonical ORF1p/ORF2p/L1-RNA-containing RNP, and we describe a nuclear population containing ORF2p, but lacking ORF1p, which likely contains host factors participating in template-primed reverse transcription.

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