title: hnRNPL and nucleolin bind LINE-1 RNA and function as host factors to modulate retrotransposition.
abstract: Long INterspersed Element one (LINE-1, or L1), is a widely distributed, autonomous retrotransposon in mammalian genomes. During retrotransposition, L1 RNA functions first as a dicistronic mRNA and then as a template for cDNA synthesis. Previously, we defined internal ribosome entry sequences (IRESs) upstream of both ORFs (ORF1 and ORF2) in the dicistronic mRNA encoded by mouse L1. Here, RNA affinity chromatography was used to isolate cellular proteins that bind these regions of L1 RNA. Four proteins, the heterogeneous nuclear ribonucleoproteins (hnRNPs) R, Q and L, and nucleolin (NCL), appeared to interact specifically with the ORF2 IRES. These were depleted from HeLa cells to examine their effects on L1 IRES-mediated translation and L1 retrotransposition. NCL knockdown specifically reduced the ORF2 IRES activity, L1 and L1-assisted Alu retrotransposition without altering L1 RNA or protein abundance. These findings are consistent with NCL acting as an IRES trans-acting factor (ITAF) for ORF2 translation and hence a positive host factor for L1 retrotransposition. In contrast, hnRNPL knockdown dramatically increased L1 retrotransposition as well as L1 RNA and ORF1 protein, indicating that this cellular protein normally interferes with retrotransposition. Thus, hnRNPL joins a small, but growing list of cellular proteins that are potent negative regulators of L1 retrotransposition.
authors.affiliation: Department of Cell and Developmental Biology, University of Colorado School of Medicine, PO Box 6511, MS 8108, Aurora, CO 80045, USA. sandy.martin@ucdenver.edu
authors.forename: Suresh
authors.initials: S
authors.lastname: Peddigari
authors.forename: Patrick Wai-Lun
authors.initials: PW
authors.lastname: Li
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Rabe
authors.forename: Sandra L
authors.initials: SL
authors.lastname: Martin
mesh_terms: D000818:Animals; D001665:Binding Sites; D004720:Endonucleases / Q000235:genetics; D006367:HeLa Cells; D034664:Heterogeneous-Nuclear Ribonucleoprotein L / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020084:Long Interspersed Nucleotide Elements; D051379:Mice; D010750:Phosphoproteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000235:genetics; D012261:Ribonucleoproteins / Q000235:genetics; D000097796:Nucleolin
chemical_list: D004720:Endonucleases; D034664:Heterogeneous-Nuclear Ribonucleoprotein L; D010750:Phosphoproteins; D012313:RNA; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D012194:RNA-Directed DNA Polymerase; D012261:Ribonucleoproteins; D000097796:Nucleolin; C561286:ECAT11 protein, mouse; C494023:L1 ORF1 protein, human; C494024:L1 ORF2 protein, human