Retrotransposon Ty1 RNA contains a 5'-terminal long-range pseudoknot required for efficient reverse transcription

Qing Huang, Katarzyna J. Purzycka, Sabrina Lusvarghi, Donghui Li, Stuart F.J. Legrice, Jef D. Boeke

Research output: Contribution to journalArticlepeer-review

Abstract

Ty1 retrotransposon RNA has the potential to fold into a variety of distinct structures, mutation of which affects retrotransposition frequencies. We show here that one potential functional structure is located at the 5' end of the genome and can assume a pseudoknot conformation. Chemoenzymatic probing of wild-type and mutant mini-Ty1 RNAs supports the existence of such a structure, while molecular genetic analyses show that mutations disrupting pseudoknot formation interfere with retrotransposition, indicating that it provides a critical biological function. These defects are enhanced at higher temperatures. When these mutants are combined with compensatory changes, retrotransposition is restored, consistent with pseudoknot architecture. Analyses of mutants suggest a defect in Ty1 reverse transcription. Collectively, our data allow modeling of a three-dimensional structure for this novel critical cis-acting signal of the Ty1 genome.

Original languageEnglish (US)
Pages (from-to)320-322
Number of pages3
JournalRNA
Volume19
Issue number3
DOIs
StatePublished - Mar 2013

Keywords

  • Pseudoknot
  • RNA structure
  • Retrotransposon
  • Reverse transcription
  • SHAPE

ASJC Scopus subject areas

  • Molecular Biology

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