LINE-1 ORF1 protein enhances Alu SINE retrotransposition.
Retroelements have contributed over one third of the human genome mass. The currently active LINE-1 (L1) codes for two proteins (ORF1p and ORF2p), both strictly required for retrotransposition. In contrast, the non-coding parasitic SINE (Alu) only appears to need the L1 ORF2p for its own amplification. This requirement was previously determined using a tissue culture assay system in human cells (HeLa). Because HeLa are likely to express functional L1 proteins, it is possible that low levels of endogenous ORF1p are necessary for the observed tagged Alu mobilization. By individually expressing ORF1 and ORF2 proteins from both human (L1RP and LRE3) and rodent (L1A102 and L1spa) L1 sources, we demonstrate that increasing amounts of ORF1 expressing vector enhances tagged Alu mobilization in HeLa cells. In addition, using chicken fibroblast cells as an alternate cell culture source, we confirmed that ORF1p is not strictly required for Alu mobilization in our assay. Supporting our observations in HeLa cells, we find that tagged Alu retrotransposition is improved by supplementation of ORF1p in the cultured chicken cells. We postulate that L1 ORF1p plays either a direct or indirect role in enhancing the interaction between the Alu RNA and the required factors needed for its retrotransposition.
Wallace
Nicholas
N
Tulane Cancer Center SL-66, Deparment of Epidemiology, Tulane University Health Sciences Center, 1430 Tulane Ave., New Orleans, LA 70112, USA.
Wagstaff
Bradley J
BJ
Deininger
Prescott L
PL
Roy-Engel
Astrid M
AM
D020087:Alu Elements; D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D020084:Long Interspersed Nucleotide Elements; D051379:Mice; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D016324:Sequence Tagged Sites
C531245:ORF1 protein, human; C531246:ORF2 protein, human; D011506:Proteins; D012194:RNA-Directed DNA Polymerase; D004720:Endonucleases