title: The Zinc-Finger Antiviral Protein ZAP Inhibits LINE and Alu Retrotransposition.
abstract: Long INterspersed Element-1 (LINE-1 or L1) is the only active autonomous retrotransposon in the human genome. To investigate the interplay between the L1 retrotransposition machinery and the host cell, we used co-immunoprecipitation in conjunction with liquid chromatography and tandem mass spectrometry to identify cellular proteins that interact with the L1 first open reading frame-encoded protein, ORF1p. We identified 39 ORF1p-interacting candidate proteins including the zinc-finger antiviral protein (ZAP or ZC3HAV1). Here we show that the interaction between ZAP and ORF1p requires RNA and that ZAP overexpression in HeLa cells inhibits the retrotransposition of engineered human L1 and Alu elements, an engineered mouse L1, and an engineered zebrafish LINE-2 element. Consistently, siRNA-mediated depletion of endogenous ZAP in HeLa cells led to a ~2-fold increase in human L1 retrotransposition. Fluorescence microscopy in cultured human cells demonstrated that ZAP co-localizes with L1 RNA, ORF1p, and stress granule associated proteins in cytoplasmic foci. Finally, molecular genetic and biochemical analyses indicate that ZAP reduces the accumulation of full-length L1 RNA and the L1-encoded proteins, yielding mechanistic insight about how ZAP may inhibit L1 retrotransposition. Together, these data suggest that ZAP inhibits the retrotransposition of LINE and Alu elements.
authors.affiliation: Cellular and Molecular Biology Graduate Program, University of Michigan, Ann Arbor, Michigan, United States of America.
authors.forename: John B
authors.initials: JB
authors.lastname: Moldovan
authors.affiliation: Cellular and Molecular Biology Graduate Program, University of Michigan, Ann Arbor, Michigan, United States of America; Departments of Human Genetics and Internal Medicine, University of Michigan, Ann Arbor, Michigan, United States of America; Howard Hughes Medical Institute, University of Michigan, Ann Arbor, Michigan, United States of America.
authors.forename: John V
authors.initials: JV
authors.lastname: Moran
mesh_terms: D020087:Alu Elements; D000818:Animals; D002853:Chromatography, Liquid; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D020084:Long Interspersed Nucleotide Elements / Q000235:genetics*; D051379:Mice; D016366:Open Reading Frames; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018626:Retroelements; D053719:Tandem Mass Spectrometry; D015027:Zebrafish
chemical_list: D016601:RNA-Binding Proteins; D018626:Retroelements; C527496:ZC3HAV1 protein, human