title: Enrichment of processed pseudogene transcripts in L1-ribonucleoprotein particles.
abstract: Long INterspersed Elements (LINE-1s, L1s) are responsible for over one million retrotransposon insertions and 8000 processed pseudogenes (PPs) in the human genome. An active L1 encodes two proteins (ORF1p and ORF2p) that bind with L1 RNA and form L1-ribonucleoprotein particles (RNPs). Although it is believed that the RNA-binding property of ORF1p is critical to recruit other mobile RNAs to the RNP, the identity of recruited RNAs is largely unknown. Here, we used crosslinking and immunoprecipitation followed by deep sequencing to identify RNA components of L1-RNPs. Our results show that in addition to retrotransposed RNAs [L1, Alu and SINE-VNTR-Alu (SVA)], L1-RNPs are enriched with cellular mRNAs, which have PPs in the human genome. Using purified L1-RNPs, we show that PP-source RNAs preferentially serve as ORF2p templates in a reverse transcriptase assay. In addition, we find that exogenous ORF2p binds endogenous ORF1p, allowing reverse transcription of the same PP-source RNAs. These data demonstrate that interaction of a cellular RNA with the L1-RNP is an inside track to PP formation.
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine and Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Prabhat K
authors.initials: PK
authors.lastname: Mandal
authors.forename: Adam D
authors.initials: AD
authors.lastname: Ewing
authors.forename: Dustin C
authors.initials: DC
authors.lastname: Hancks
authors.forename: Haig H
authors.initials: HH
authors.lastname: Kazazian
mesh_terms: D015870:Gene Expression; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D020084:Long Interspersed Nucleotide Elements / Q000235:genetics*; D016366:Open Reading Frames; D011544:Pseudogenes; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism; D018626:Retroelements; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D016601:RNA-Binding Proteins; D018626:Retroelements; D012261:Ribonucleoproteins; D012313:RNA; D012194:RNA-Directed DNA Polymerase