A 3' Poly(A) Tract Is Required for LINE-1 Retrotransposition

Aurélien J Doucet; Jeremy E Wilusz; Tomoichiro Miyoshi; Ying Liu; John V Moran

doi:10.1016/j.molcel.2015.10.012

A 3' Poly(A) Tract Is Required for LINE-1 Retrotransposition

Mol Cell. 2015 Dec 3;60(5):728-741. doi: 10.1016/j.molcel.2015.10.012. Epub 2015 Nov 12.

Authors

Aurélien J Doucet¹, Jeremy E Wilusz², Tomoichiro Miyoshi³, Ying Liu⁴, John V Moran⁵

Affiliations

¹ Department of Human Genetics, University of Michigan Medical School, Ann Arbor, MI 48109-5618, USA. Electronic address: adoucet@umich.edu.
² Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
³ Department of Human Genetics, University of Michigan Medical School, Ann Arbor, MI 48109-5618, USA.
⁴ Howard Hughes Medical Institute, University of Michigan Medical School, Ann Arbor, MI 48109-5618, USA.
⁵ Department of Human Genetics, University of Michigan Medical School, Ann Arbor, MI 48109-5618, USA; Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48109-5618, USA; Howard Hughes Medical Institute, University of Michigan Medical School, Ann Arbor, MI 48109-5618, USA. Electronic address: moranj@umich.edu.

Abstract

L1 retrotransposons express proteins (ORF1p and ORF2p) that preferentially mobilize their encoding RNA in cis, but they also can mobilize Alu RNA and, more rarely, cellular mRNAs in trans. Although these RNAs differ in sequence, each ends in a 3' polyadenosine (poly(A)) tract. Here, we replace the L1 polyadenylation signal with sequences derived from a non-polyadenylated long non-coding RNA (MALAT1), which can form a stabilizing triple helix at the 3' end of an RNA. L1/MALAT RNAs accumulate in cells, lack poly(A) tails, and are translated; however, they cannot retrotranspose in cis. Remarkably, the addition of a 16 or 40 base poly(A) tract downstream of the L1/MALAT triple helix restores retrotransposition in cis. The presence of a poly(A) tract also allows ORF2p to bind and mobilize RNAs in trans. Thus, a 3' poly(A) tract is critical for the retrotransposition of sequences that comprise approximately one billion base pairs of human DNA.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Endonucleases / genetics
HeLa Cells
Humans
Long Interspersed Nucleotide Elements*
Poly A / metabolism*
RNA, Long Noncoding / metabolism
RNA, Messenger / chemistry*
RNA, Messenger / genetics
RNA-Directed DNA Polymerase / genetics

Substances

MALAT1 long non-coding RNA, human
ORF2 protein, human
RNA, Long Noncoding
RNA, Messenger
Poly A
RNA-Directed DNA Polymerase
Endonucleases

Abstract

Publication types

MeSH terms

Substances

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