Life cycle of yeast dsRNA viruses uncovered by advanced fluorescent microscopy

Duration of the project

01.07.2019. - 03.10.2019.

Countries and institutions involved in the project

flag-LT
Vilnius University
flag-DE
University of Heidelberg

Aim of the project

Saccharomyces yeast possesses 23% genes homologous to human counterparts and is highly relevant model system for functional studies of Homo sapiens cells. Double-stranded RNA viruses of Saccharomyces cerevisiae are remarkable for lack of extracellular stage. They are widespread in numerous strains, including BY4741 – host of various genome-wide libraries. The capability of virus – benign component of the cell –- to significantly alter the transcriptomic landscape of yeast has been demonstrated recently. Thus, all transcriptomic studies conducted in dsRNA virus-possessing yeast should account for the virus integrity; however, few if any of such studies do report doing so. Factors affecting intrinsic virus might execute profound changes on transcriptome, potentially compromising results of unrelated studies.

To address the life cycle of L-A-1 virus of BY4741 strain, we cloned, sequenced and expressed genome of virus. Capsid protein was tagged to convey optical traceability of viral protein, competent for virion formation. Expression of proteins from inducible promoter allows time-resolved tracking, while use of compartmentalization markers provides the spatial resolution. The ultimate goal is observation of complete viral cycle from synthesis start to formation of mature virions, never reported before for dsRNA virus and of high interest due to ultimate integration of virus into yeast metabolism.

Main activities of the project

  1. Initiation of the Project will be accomplished at the home institution (Life Sciences Center, Vilnius). The construction and verification of corresponding yeast strains along with functional analysis for virus maintenance will be performed.
  2. Analysis of localization of viral proteins and compartmentalization markers in yeast cells will be performed by variety of fluorescent microscopy techniques at University of Heidelberg, Advanced Biological Screening Facility.

Target group and number of persons involved

Direct: members of Prof. Saulius Serva and Dr. Holger Erfle research groups (up to 15).

Indirect: employees of Life Science Center (Vilnius, Lithuania) and BioQuant along with Department of Infectious Diseases/Virology (Heidelberg, Germany) – more than 500. Also, whole scientific community would benefit upon the dissemination of research results (publication is planned).

Public events

Public seminar in University of Heidelberg is planned to be held on October 11, 2019. Speakers: Dr. Saulius Serva, Dr. Manuel Gunkel.