Article (Scientific journals)
The non-autonomous retrotransposon SVA is trans-mobilized by the human LINE-1 protein machinery
Raiz, J.; Damert, A.; Chira, S. et al.
2012In Nucleic Acids Research, 40 (4), p. 1666-1683
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Keywords :
DNA content; HeLa cell; SVA; Alu Elements; Endonucleases; Genes, Reporter; Genetic Engineering; HeLa Cells; Humans; Retroelements; Ribonucleoproteins; RNA-Directed DNA Polymerase; Transduction, Genetic; Hominidae
Abstract :
[en] 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. © 2012 The Author(s).
Disciplines :
Biochemistry, biophysics & molecular biology
Identifiers :
eid=2-s2.0-84857888415
Author, co-author :
Raiz, J.;  Section PR2/Retroelements, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany
Damert, A.;  Section PR2/Retroelements, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany, Institute for Interdisciplinary Research in Bio-Nano-Sciences, Molecular Biology Center, Babes-Bolyai-University, Cluj-Napoca, Treboniu Laurean Street 42, RO-400271 Cluj-Napoca, Romania
Chira, S.;  Institute for Interdisciplinary Research in Bio-Nano-Sciences, Molecular Biology Center, Babes-Bolyai-University, Cluj-Napoca, Treboniu Laurean Street 42, RO-400271 Cluj-Napoca, Romania
Held, U.;  Section PR2/Retroelements, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany, Division of Medical Biotechnology, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany
Klawitter, S.;  Division of Medical Biotechnology, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany
Hamdorf, Matthias ;  University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit
Löwer, J.;  Section PR2/Retroelements, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany
Strätling, W. H.;  Institut für Biochemie und Molekularbiologie, Universitätsklinikum Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany
Löwer, R.;  Section PR2/Retroelements, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany
Schumann, G. G.;  Section PR2/Retroelements, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany, Division of Medical Biotechnology, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany
External co-authors :
yes
Language :
English
Title :
The non-autonomous retrotransposon SVA is trans-mobilized by the human LINE-1 protein machinery
Publication date :
2012
Journal title :
Nucleic Acids Research
ISSN :
0305-1048
Volume :
40
Issue :
4
Pages :
1666-1683
Peer reviewed :
Peer reviewed
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