Abstract

The yeasts Torulaspora delbrueckii (Td) and Saccharomyces cerevisiae (Sc) may show a killer phenotype that is encoded in dsRNA M viruses (V-M), which require the helper activity of another dsRNA virus (V-LA or V-LBC) for replication. Recently, two TdV-LBCbarr genomes, which share sequence identity with ScV-LBC counterparts, were characterized by high-throughput sequencing (HTS). They also share some similar characteristics with Sc-LA viruses. This may explain why TdV-LBCbarr has helper capability to maintain M viruses, whereas ScV-LBC does not. We here analyze two stretches with low sequence identity (LIS I and LIS II) that were found in TdV-LBCbarr Gag-Pol proteins when comparing with the homologous regions of ScV-LBC. These stretches may result from successive nucleotide insertions or deletions (indels) that allow compensatory frameshift events required to maintain specific functions of the RNA-polymerase, while modifying other functions such as the ability to bind V-M (+)RNA for packaging. The presence of an additional frameshifting site in LIS I may ensure the synthesis of a certain amount of RNA-polymerase until the new compensatory indel appears. Additional 5′- and 3′-extra sequences were found beyond V-LBC canonical genomes. Most extra sequences showed high identity to some stretches of the canonical genomes and can form stem-loop structures. Further, the 3′-extra sequence of two ScV-LBC genomes contains rRNA stretches. The origin and possible functions of these extra sequences are here discussed.

Highlights

  • Killer yeasts produce protein toxins that are lethal to sensitive yeasts

  • Virus that may coexist with virus with a large-size genome (V-LA) and virus with a medium-size genome (V-M) in the cytoplasm of S. cerevisiae; no helper activity is known for V-LBC in this yeast species [4,5,6,7,8]

  • In addition to the conserved slippery site found in all known LBC viruses upstream from the Gag ORF stop codon, TdV-LBCbarr genomes have a second putative in-frame translation re-initiation codon (“2151GGGGAGATGA2160”) located downstream from Gag-ORF and upstream from Pol domain

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Summary

Introduction

The synthesis and secretion of killer toxins by Torulaspora delbrueckii (Td) and Saccharomyces cerevisiae (Sc) requires the presence of at least two cytoplasmic dsRNA viruses that are members of the family Totiviridae. A new LBC virus has recently been found in T. delbrueckii (TdV-LBCbarr2) that may act as a helper for two M viruses in the same yeast strain: TdV-Mbarr and ScV-M1 [9]. These viruses are inherited in the cytoplasm from mother yeast to daughter bud and transferred horizontally between different yeasts by mating or heterokaryon formation [10].

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