Completion of LINE integration involves an open '4-way' branched DNA intermediate

Nucleic Acids Research
Brijesh B KhadgiShawn M Christensen

Abstract

Long Interspersed Elements (LINEs), also known as non-LTR retrotransposons, encode a multifunctional protein that reverse transcribes its mRNA into DNA at the site of insertion by target primed reverse transcription. The second half of the integration reaction remains very poorly understood. Second-strand DNA cleavage and second-strand DNA synthesis were investigated in vitro using purified components from a site-specific restriction-like endonuclease (RLE) bearing LINE. DNA structure was shown to be a critical component of second-strand DNA cleavage. A hitherto unknown and unexplored integration intermediate, an open '4-way' DNA junction, was recognized by the element protein and cleaved in a Holliday junction resolvase-like reaction. Cleavage of the 4-way junction resulted in a natural primer-template pairing used for second-strand DNA synthesis. A new model for RLE LINE integration is presented.

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Citations

Dec 10, 2019·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Dragomira N MarkovaEsther Betrán
Sep 23, 2021·Nucleic Acids Research·Getong LiuAlastair I H Murchie

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Methods Mentioned

BETA
electrophoresis
electrophoretic mobility shift assay

Software Mentioned

FIJI

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