Yeast screens for host factors in positive-strand RNA virus replication based on a library of temperature-sensitive mutants

Muhammad Shah Nawaz-ul-Rehman, K. Reddisiva Prasanth, Jannine Baker, Peter D. Nagy

Research output: Contribution to journalArticlepeer-review

43 Scopus citations


RNA viruses exploit host cells by altering cellular pathways, recruiting host factors, remodeling intracellular membranes and escaping host antiviral responses. Model hosts, such as Saccharomyces cerevisiae (yeast), are valuable to identify host factors involved in viral RNA replication. The many advantages of using yeast include the availability of various yeast mutant libraries, such as (i) single gene-deletion library; (ii) the essential gene library (yTHC); and (iii) the yeast ORF over-expression library. Here, we have used a novel temperature-sensitive (ts) mutant library of essential yeast genes to identify 118 host proteins affecting replication of Tomato bushy stunt virus, in yeast model host. Testing 787 ts mutants led to the identification of host factors, of which 72 proteins facilitated TBSV replication in yeast and 46 proteins were inhibitory. Altogether, ∼85% of the identified proteins are novel host factors affecting tombusvirus replication. The ts mutant library screen also led to the identification of 17 essential genes, which have been documented before, thus confirming the importance of these genomic screens. Overall, we show the power of ts mutant library in identification of host factors for RNA virus replication.

Original languageEnglish
Pages (from-to)207-216
Number of pages10
Issue number2
StatePublished - Feb 2013

Bibliographical note

Funding Information:
The authors thank Dr. Charlie Boone (Banting and Best Department of Medical Research and Department of Molecular Genetics, The Donnelly Centre, University of Toronto, Toronto, Ontario, Canada) for providing the yeast ts-mutant library. We thank Drs. Zhijian Li (Univ. Toronto) and Judit Pogany (UK, Lexington) for very helpful suggestions. This work was supported by NIAID ( 5R21AI079457-02 ) and the Kentucky Science Foundation .


  • High throughput screen
  • Host factors
  • RNA virus
  • Replication
  • Temperature-sensitive
  • Tomato bushy stunt virus
  • Virus - host interaction
  • Yeast

ASJC Scopus subject areas

  • Molecular Biology
  • General Biochemistry, Genetics and Molecular Biology


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