Multiple molecular determinants for retrotransposition in a primer tRNA

Jill B. Keeney, Karen B. Chapman, Vit Lauermann, Daniel F. Voytas, Stefan U. Åström, Ulrich Von Pawel-Rammingen, Anders Byström, Jef D. Boeke

Research output: Contribution to journalArticle

Abstract

Retroviruses and long terminal repeat-containing retroelements use host- encoded tRNAs as primers for the synthesis of minus strong-stop DNA, the first intermediate in reverse transcription of the retroelement RNA. Usually, one or more specific tRNAs, including the primer, are selected and packaged within the virion. The reverse transcriptase (RT) interacts with the primer tRNA and initiates DNA synthesis. The structural and sequence features of primer tRNAs important for these specific interactions are poorly understood. We have developed a genetic assay in which mutants of tRNA(i)/(Met), the primer for the Ty1 retrotransposon of Saccharomyces cerevisiae, can be tested for the ability to serve as primers in the reverse transcription process. This system allows any tRNA mutant to be tested, regardless of its ability to function in the initiation of protein synthesis. We find that mutations in the TΨC loop and the acceptor stem regions of the tRNA(i)/(Met) affect transposition most severely. Conversely, mutations in the anticodon region have only minimal effects on transposition. Further study of the acceptor stem and other mutants demonstrates that complementarity to the element primer binding site is a necessary but not sufficient requirement for effective tRNA priming. Finally, we have used interspecies hybrid initiator tRNA molecules to implicate nucleotides in the D arm as additional recognition determinants. Ty3 and Ty1, two very distantly related retrotransposons, require similar molecular determinants in this primer tRNA for transposition.

Original languageEnglish (US)
Pages (from-to)217-226
Number of pages10
JournalMolecular and Cellular Biology
Volume15
Issue number1
StatePublished - Jan 1995

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Transfer RNA
Retroelements
RNA, Transfer, Met
Reverse Transcription
Anticodon
Mutation
Terminal Repeat Sequences
RNA-Directed DNA Polymerase
Retroviridae
Virion
Saccharomyces cerevisiae
Nucleotides
Binding Sites
RNA
DNA
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Keeney, J. B., Chapman, K. B., Lauermann, V., Voytas, D. F., Åström, S. U., Von Pawel-Rammingen, U., ... Boeke, J. D. (1995). Multiple molecular determinants for retrotransposition in a primer tRNA. Molecular and Cellular Biology, 15(1), 217-226.

Multiple molecular determinants for retrotransposition in a primer tRNA. / Keeney, Jill B.; Chapman, Karen B.; Lauermann, Vit; Voytas, Daniel F.; Åström, Stefan U.; Von Pawel-Rammingen, Ulrich; Byström, Anders; Boeke, Jef D.

In: Molecular and Cellular Biology, Vol. 15, No. 1, 01.1995, p. 217-226.

Research output: Contribution to journalArticle

Keeney, JB, Chapman, KB, Lauermann, V, Voytas, DF, Åström, SU, Von Pawel-Rammingen, U, Byström, A & Boeke, JD 1995, 'Multiple molecular determinants for retrotransposition in a primer tRNA', Molecular and Cellular Biology, vol. 15, no. 1, pp. 217-226.
Keeney JB, Chapman KB, Lauermann V, Voytas DF, Åström SU, Von Pawel-Rammingen U et al. Multiple molecular determinants for retrotransposition in a primer tRNA. Molecular and Cellular Biology. 1995 Jan;15(1):217-226.
Keeney, Jill B. ; Chapman, Karen B. ; Lauermann, Vit ; Voytas, Daniel F. ; Åström, Stefan U. ; Von Pawel-Rammingen, Ulrich ; Byström, Anders ; Boeke, Jef D. / Multiple molecular determinants for retrotransposition in a primer tRNA. In: Molecular and Cellular Biology. 1995 ; Vol. 15, No. 1. pp. 217-226.
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