title: The non-autonomous retrotransposon SVA is trans-mobilized by the human LINE-1 protein machinery.
abstract: SINE-VNTR-Alu (SVA) elements are non-autonomous, hominid-specific non-LTR retrotransposons and distinguished by their organization as composite mobile elements. They represent the evolutionarily youngest, currently active family of human non-LTR retrotransposons, and sporadically generate disease-causing insertions. Since preexisting, genomic SVA sequences are characterized by structural hallmarks of Long Interspersed Elements 1 (LINE-1, L1)-mediated retrotransposition, it has been hypothesized for several years that SVA elements are mobilized by the L1 protein machinery in trans. To test this hypothesis, we developed an SVA retrotransposition reporter assay in cell culture using three different human-specific SVA reporter elements. We demonstrate that SVA elements are mobilized in HeLa cells only in the presence of both L1-encoded proteins, ORF1p and ORF2p. SVA trans-mobilization rates exceeded pseudogene formation frequencies by 12- to 300-fold in HeLa-HA cells, indicating that SVA elements represent a preferred substrate for L1 proteins. Acquisition of an AluSp element increased the trans-mobilization frequency of the SVA reporter element by ~25-fold. Deletion of (CCCTCT)(n) repeats and Alu-like region of a canonical SVA reporter element caused significant attenuation of the SVA trans-mobilization rate. SVA de novo insertions were predominantly full-length, occurred preferentially in G+C-rich regions, and displayed all features of L1-mediated retrotransposition which are also observed in preexisting genomic SVA insertions.
authors: Section PR2/Retroelements, Division of Medical Biotechnology, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany.
authors: Julija
authors: J
authors: Raiz
authors: Annette
authors: A
authors: Damert
authors: Sergiu
authors: S
authors: Chira
authors: Ulrike
authors: U
authors: Held
authors: Sabine
authors: S
authors: Klawitter
authors: Matthias
authors: M
authors: Hamdorf
authors: Johannes
authors: J
authors: Löwer
authors: Wolf H
authors: WH
authors: Strätling
authors: Roswitha
authors: R
authors: Löwer
authors: Gerald G
authors: GG
authors: Schumann
mesh_terms: D020087:Alu Elements; D004720:Endonucleases / Q000378:metabolism*; D017930:Genes, Reporter; D005818:Genetic Engineering; D006367:HeLa Cells; D006801:Humans; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D018626:Retroelements; D012261:Ribonucleoproteins / Q000378:metabolism*; D014161:Transduction, Genetic
chemical_list: C494023:L1 ORF1 protein, human; C494024:L1 ORF2 protein, human; D018626:Retroelements; D012261:Ribonucleoproteins; D012194:RNA-Directed DNA Polymerase; D004720:Endonucleases