Cell division promotes efficient retrotransposition in a stable L1 reporter cell line

Xie, Yi and Mátés, Lajos and Ivics, Zoltán and Izsvák, Zsuzsanna and Martin, Sandra L. and An, Wenfeng (2013) Cell division promotes efficient retrotransposition in a stable L1 reporter cell line. MOBILE DNA, 4. ISSN 1759-8753


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Background: Long interspersed element type one (L1) actively modifies the human genome by inserting new copies of itself. This process, termed retrotransposition, requires the formation of an L1 ribonucleoprotein (RNP) complex, which must enter the nucleus before retrotransposition can proceed. Thus, the nuclear import of L1 RNP presents an opportunity for cells to regulate L1 retrotransposition post-translationally. The effect of cell division on L1 retrotransposition has been investigated by two previous studies, which observed varied degrees of inhibition in retrotransposition when primary cell strains or cancer cell lines were experimentally arrested in different stages of the cell cycle. However, seemingly divergent conclusions were reached. The role of cell division on retrotransposition remains highly debated. Findings: To monitor both L1 expression and retrotransposition quantitatively, we developed a stable dual-luciferase L1 reporter cell line, in which a bi-directional tetracycline-inducible promoter drives the expression of both a firefly luciferase-tagged L1 element and a Renilla luciferase, the latter indicative of the level of promoter induction. We observed an additional 10-fold reduction in retrotransposition in cell-cycle arrested cells even after retrotransposition had been normalized to Renilla luciferase or L1 ORF1 protein levels. In synchronized cells, cells undergoing two mitoses showed 2.6-fold higher retrotransposition than those undergoing one mitosis although L1 expression was induced for the same amount of time. Conclusions: Our data provide additional support for an important role of cell division in retrotransposition and argue that restricting the accessibility of L1 RNP to nuclear DNA could be a post-translational regulatory mechanism for retrotransposition.

Item Type: Article
Uncontrolled Keywords: FETAL; EVOLUTION; RNA; PROTEIN; EXPRESSION; OPEN READING FRAME-1; stable cell line; TRANSCRIPTION; Tetracycline-inducible promoter; Nuclear import; Non-LTR retrotransposon; LINE-1; Dual-luciferase assay; Cell Division; Cell-cycle synchronization; CELL-CYCLE ARREST
Subjects: Q Science / természettudomány > QH Natural history / természetrajz > QH301 Biology / biológia
Depositing User: MTMT SWORD
Date Deposited: 01 Oct 2013 10:39
Last Modified: 01 Oct 2013 10:39

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