Epigenetic silencing of engineered L1 retrotransposition events in human embryonic carcinoma cells

• Type of Material: Article
• Authors: Jose L. Garcia-Perez, University of Michigan; Maria Morell, University of Granada; Joshua O. Scheys, University of Michigan; Deanna A. Kulpa, Emory University; Santiago Morell, University of Granada; Christoph C. Carter, University of Michigan; Gary D. Hammer, University of Michigan; Kathleen L. Collins, University of Michigan; K. Sue O'Shea, University of Michigan; Pablo Menendez, University of Granada; John V. Moran, University of Michigan
• Language: English
• Date: 2010-08-05
• Publisher: Nature Publishing Group
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: ©2010 Macmillan Publishers Limited. All rights reserved
• Final Published Version (URL): http://dx.doi.org/10.1038/nature09209
• Title of Journal or Parent Work: Nature
• ISSN: 0028-0836
• Volume: 466
• Issue: 7307
• Start Page: 769
• End Page: 773
• Grant/Funding Information: J.V.M. is supported by the National Institutes of Health (NIH) (GM060518 and GM082970) and the Howard Hughes Medical Institute. J.L.G.-P. is supported by the Instituto de Salud Carlos III - Consejeria de Salud Junta de Andalucia (ISCIII-CSJA) (EMER07/056), by a Marie Curie International Reintegration Grant action (FP7-PEOPLE-2007-4-3-IRG), by CICE (P09-CTS-4980) and Proyectos en Salud (PI0002/2009) from Junta de Andalucia (Spain) and through the Spanish Ministry of Health (FIS PI08171 and Miguel Servet CP07/00065).; J.O.S. is supported by a Cellular and Molecular Approaches to Systems and Integrative Biology Training Grant (T32-GM08322).; K.S.O’S. is supported by the NIH (NS-048187 and GM-069985).; K.L.C. is supported by the Burroughs Wellcome Foundation and by an NIH Research Project Grant (R01) (AI051198).; M.M. is supported by the ISCIII-CSJA (EMER07/056). P.M. is supported by the Spanish Ministry of Science and Innovation MICINN-PLANE (PLE-2009-0111), by CICE (P08-CTS-3678) from Junta de Andalucia (Spain) and by the Spanish Ministry of Health (FIS PI070026).; G.D.H is supported by a National Institute of Diabetes and Digestive and Kidney Diseases NIH R01 (DK62027).; We defrayed the costs of DNA sequencing in part with the University of Michigan's Cancer Center Support Grant (NIH 5 P30 CA46592).; D.A.K. is supported by The Irvington Institute Fellowship Program of the Cancer Research Institute.; S.M. is supported by a CICE (P08-CTS-3678) scholarship from Junta de Andalucia, Spain. C.C.C. is supported by a Rackham Predoctoral Fellowship from the University of Michigan.
• Keywords: MOUSE; Science & Technology - Other Topics; VIRUS; PATTERNS; PROTEIN; EXPRESSION; Multidisciplinary Sciences; Science & Technology; STEM-CELLS; SOMATIC MOSAICISM; LINE-1 RETROTRANSPOSITION
• Research Categories: Biology, General; Biology, Molecular

Abstract

Long interspersed element-1 (LINE-1 or L1) retrotransposition continues to affect human genome evolution 1,2 . L1s can retrotranspose in the germline, during early development and in select somatic cells 3-8 ; however, the host response to L1 retrotransposition remains largely unexplored. Here we show that reporter genes introduced into the genome of various human embryonic carcinoma-derived cell lines (ECs) by L1 retrotransposition are rapidly and efficiently silenced either during or immediately after their integration. Treating ECs with histone deacetylase inhibitors rapidly reverses this silencing, and chromatin immunoprecipitation experiments revealed that reactivation of the reporter gene was correlated with changes in chromatin status at the L1 integration site. Under our assay conditions, rapid silencing was also observed when reporter genes were delivered into ECs by mouse L1s and a zebrafish LINE-2 element, but not when similar reporter genes were delivered into ECs by Moloney murine leukaemia virus or human immunodeficiency virus, suggesting that these integration events are silenced by distinct mechanisms. Finally, we demonstrate that subjecting ECs to culture conditions that promote differentiation attenuates the silencing of reporter genes delivered by L1 retrotransposition, but that differentiation, in itself, is not sufficient to reactivate previously silenced reporter genes. Thus, our data indicate that ECs differ from many differentiated cells in their ability to silence reporter genes delivered by L1 retrotransposition. © 2010 Macmillan Publishers Limited. All rights reserved.