Abstract

The long-terminal repeat retrotransposon Ty1 is the most abundant mobile genetic element in many Saccharomyces cerevisiae isolates. Ty1 retrotransposons contribute to the genetic diversity of host cells, but they can also act as an insertional mutagen and cause genetic instability. Interestingly, retrotransposition occurs at a low level despite a high level of Ty1 RNA, even though S. cerevisiae lacks the intrinsic defense mechanisms that other eukaryotes use to prevent transposon movement. p22 is a recently discovered Ty1 protein that inhibits retrotransposition in a dose-dependent manner. p22 is a truncated form of Gag encoded by internally initiated Ty1i RNA that contains two closely-spaced AUG codons. Mutations of either AUG codon compromise p22 translation. We found that both AUG codons were utilized and that translation efficiency depended on the Ty1i RNA structure. Structural features that stimulated p22 translation were context dependent and present only in Ty1i RNA. Destabilization of the 5′ untranslated region (5′ UTR) of Ty1i RNA decreased the p22 level, both in vitro and in vivo. Our data suggest that protein factors such as Gag could contribute to the stability and translational activity of Ty1i RNA through specific interactions with structural motifs in the RNA.

Highlights

  • Ty1 is a long-terminal repeat (LTR) retrotransposon in the Pseudoviridae family and the most abundant mobile genetic element in the Saccharomyces cerevisiae reference strain [1]

  • Dimeric Ty1 RNA is present in virus-like particles (VLPs) [3] that are comprised of the capsid protein Gag and Gag-Pol; the latter being synthesized by a programmed +1 frameshift event that occurs at overlapping leucine codons in GAG and POL [4]

  • This coordinated action allows cells to rapidly adapt to their environment without the need of de novo mRNA synthesis and transport from the nucleus to the cytoplasm [50]

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Summary

Introduction

Ty1 is a long-terminal repeat (LTR) retrotransposon in the Pseudoviridae family and the most abundant mobile genetic element in the Saccharomyces cerevisiae reference strain [1]. GAG and POL genes bracketed by LTRs and proliferates in the yeast genome by integrating new copies through an RNA-mediated mechanism [2]. POL encodes protease (PR), reverse transcriptase (RT) and integrase (IN), which are required for protein maturation, reverse transcription and integration, respectively. Gag is a VLP structural component and is expressed as a 441-amino acid precursor (p49) that undergoes a C-terminal cleavage by PR to produce the mature 401-residue protein (p45). Ty1 Gag binds RNA in vitro [5,6] and serves as a multifunctional regulator that orchestrates retrotransposon replication [7]

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