Cis-preferential LINE-1 reverse transcriptase activity in ribonucleoprotein particles.
LINE-1 retrotransposons (L1s) constitute approximately 17% of human DNA, and their activity continues to affect genome evolution. Retrotransposition-competent human L1s encode two proteins required for their mobility (ORF1p and ORF2p); however, biochemical activities associated with ORF2p have been difficult to detect in cells. Here, we show for the first time the colocalization of L1 RNA, ORF1p and ORF2p to a putative ribonucleoprotein retrotransposition intermediate. We further demonstrate that ORF2p preferentially uses its encoding RNA as a template for reverse transcription. Thus, our data provide the first biochemical evidence supporting the cis-preferential action of the L1 reverse transcriptase.
Kulpa
Deanna A
DA
Department of Human Genetics 1241 E. Catherine St., University of Michigan Medical School, Ann Arbor, Michigan 48109-0618, USA. kulpadea@umich.edu
Moran
John V
JV
D001483:Base Sequence; D006367:HeLa Cells; D006801:Humans; D020084:Long Interspersed Nucleotide Elements / Q000235:genetics*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016366:Open Reading Frames; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D018626:Retroelements / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012261:Ribonucleoproteins / Q000378:metabolism*; D014162:Transfection
D011994:Recombinant Proteins; D018626:Retroelements; D012261:Ribonucleoproteins; D012194:RNA-Directed DNA Polymerase