Interactions between Ty1 retrotransposon RNA and the T and D regions of the tRNA(iMet) primer are required for initiation of reverse transcription in vivo.

Molecular and Cellular Biology
S FriantF X Wilhelm

Abstract

Reverse transcription of the Saccharomyces cerevisiae Ty1 retrotransposon is primed by tRNA(iMet) base paired to the primer binding site (PBS) near the 5' end of Ty1 genomic RNA. The 10-nucleotide PBS is complementary to the last 10 nucleotides of the acceptor stem of tRNA(iMet). A structural probing study of the interactions between the Ty1 RNA template and the tRNA(iMet) primer showed that besides interactions between the PBS and the 3' end of tRNA(iMet), three short regions of Ty1 RNA, named boxes 0, 1, and 2.1, interact with the T and D stems and loops of tRNA(iMet). To determine if these sequences are important for the reverse transcription pathway of the Ty1 retrotransposon, mutant Ty1 elements and tRNA(iMet) were tested for the ability to support transposition. We show that the Ty1 boxes and the complementary sequences in the T and D stems and loops of tRNA(iMet) contain bases that are critical for Ty1 retrotransposition. Disruption of 1 or 2 bp between tRNA(iMet) and box 0, 1, or 2.1 dramatically decreases the level of transposition. Compensatory mutations which restore base pairing between the primer and the template restore transposition. Analysis of the reverse transcription intermediates generated inside Ty1 virus-l...Continue Reading

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Citations

Oct 18, 2012·Virus Research·Kristina ClemensSuzanne Sandmeyer
Oct 25, 2012·Nucleic Acids Research·Katarzyna J PurzyckaDavid J Garfinkel
Dec 4, 2002·Proceedings of the National Academy of Sciences of the United States of America·Mamuka KvaratskheliaStuart F J Le Grice
Feb 6, 2004·Journal of Molecular Biology·Christian Kroun DamgaardJørgen Kjems
Aug 12, 2005·Cytogenetic and Genome Research·F-X WilhelmA Gabriel
Aug 27, 2002·The Journal of Biological Chemistry·Valerie GoldschmidtRoland Marquet
Dec 30, 2010·RNA Biology·Suzanne B Sandmeyer, Kristina A Clemens
Aug 31, 2019·RNA Biology·Julita GumnaKatarzyna Pachulska-Wieczorek
Feb 24, 2021·Nucleic Acids Research·Angelika AndrzejewskaKatarzyna Pachulska-Wieczorek
Jun 25, 2015·Microbiology Spectrum·M Joan CurcioPascale Lesage
Aug 28, 2021·International Journal of Molecular Sciences·Julita GumnaKatarzyna Pachulska-Wieczorek

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