The minimal active human SVA retrotransposon requires only the 5′-Hexamer and Alu-like domains

Dustin C. Hancks, Prabhat K. Mandal, Ling E. Cheung, Haig H. Kazazian

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21 Scopus citations

Abstract

RNA-based duplication mediated by reverse transcriptase (RT), a process termed retrotransposition, is ongoing in humans and is a source of significant inter- and perhaps intraindividual genomic variation. The long interspersed element 1 (LINE-1 or L1) ORF2 protein is the genomic source for RT activity required for mobilization of its own RNA in cis and other RNAs, such as SINE/variable-number tandem-repeat (VNTR)/Alu (SVA) elements, in trans. SVA elements are~2-kb hominid-specific noncoding RNAs that have resulted in single-gene disease in humans through insertional mutagenesis or aberrant mRNA splicing. Here, using an SVA retrotransposition cell culture assay in U2OS cells, we investigated SVA domains important in L1-mediated SVA retrotransposition. Partial- and whole-domain deletions revealed that removal of either the Alu-like or SINE-R domain in the context of a full-length SVA has little to no effect, whereas removal of the CT hexamer or the VNTR domain can result in a 75% decrease in activity. Additional experiments demonstrate that the Alu-like fragment alone can retrotranspose at low levels while the addition of the CT hexamer can enhance activity as much as 2-fold compared to that of the full-length SVA. These results suggest that no SVA domain is essential for retrotransposition in U2OS cells and that the 5= end of SVA (hexamer and Alu-like domain) is sufficient for retrotransposition.

Original languageEnglish (US)
Pages (from-to)4718-4726
Number of pages9
JournalMolecular and Cellular Biology
Volume32
Issue number22
DOIs
Publication statusPublished - Nov 1 2012

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ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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