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 journalArticle

32 Citations (Scopus)

Abstract

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 (US)
Pages (from-to)207-216
Number of pages10
JournalMethods
Volume59
Issue number2
DOIs
StatePublished - Feb 2013
Externally publishedYes

Fingerprint

RNA Viruses
Virus Replication
Viruses
Yeast
Libraries
Yeasts
RNA
Temperature
Essential Genes
Tombusvirus
Gene Library
Genes
Integration Host Factors
Intracellular Membranes
Proteins
Fungal Proteins
Gene Deletion
Viral RNA
Open Reading Frames
Antiviral Agents

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Yeast screens for host factors in positive-strand RNA virus replication based on a library of temperature-sensitive mutants. / Nawaz-ul-Rehman, Muhammad Shah; Reddisiva Prasanth, K.; Baker, Jannine; Nagy, Peter D.

In: Methods, Vol. 59, No. 2, 02.2013, p. 207-216.

Research output: Contribution to journalArticle

Nawaz-ul-Rehman, Muhammad Shah ; Reddisiva Prasanth, K. ; Baker, Jannine ; Nagy, Peter D. / Yeast screens for host factors in positive-strand RNA virus replication based on a library of temperature-sensitive mutants. In: Methods. 2013 ; Vol. 59, No. 2. pp. 207-216.
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