title: Screening a yeast library of temperature-sensitive mutants reveals a role for actin in tombusvirus RNA recombination.
abstract: Genetic recombination in RNA viruses drives the evolutionary arms race with host's antiviral strategies and recombination also facilitates adaptation of viruses to new hosts. In this paper, the authors used tombusvirus and a temperature-sensitive (ts) mutant library of yeast to identify 40 host proteins affecting viral recombination in yeast model host. Subsequent detailed analysis with two identified actin-related proteins, Act1p and Arp3p, has revealed that the wt actin network helps TBSV to maintain low level viral recombination. Pharmacological inhibition of actin in plant protoplasts confirmed the role of the actin network in tombusvirus recombination. An in vitro approach revealed the altered activity of the tombusvirus replicase in the presence of mutated Act1p. The authors show more efficient recruitment of a cellular DEAD-box helicase, which enhances tombusvirus recombination, into the membrane-bound replicase in Act1p mutant yeast. Overall, this work shows that the actin network affects tombusvirus recombination in yeast and plant cells.
authors.affiliation: Department of Plant Pathology, University of Kentucky, Plant Science Building, Lexington, KY 40546, USA.
authors.forename: K Reddisiva
authors.initials: KR
authors.lastname: Prasanth
authors.affiliation: Department of Plant Pathology, University of Kentucky, Plant Science Building, Lexington, KY 40546, USA.
authors.forename: Nikolay
authors.initials: N
authors.lastname: Kovalev
authors.affiliation: Cell Structure Laboratory, Centro Nacional de Biotecnología (CNB-CSIC), Campus de Cantoblanco, 28049 Madrid, Spain.
authors.forename: Isabel Fernández
authors.initials: IF
authors.lastname: de Castro Martín
authors.affiliation: Department of Plant Pathology, University of Kentucky, Plant Science Building, Lexington, KY 40546, USA.
authors.forename: Jannine
authors.initials: J
authors.lastname: Baker
authors.affiliation: Department of Plant Pathology, University of Kentucky, Plant Science Building, Lexington, KY 40546, USA. Electronic address: pdnagy2@uky.edu.
authors.forename: Peter D
authors.initials: PD
authors.lastname: Nagy
mesh_terms: D051378:Actin-Related Protein 3 / Q000235:genetics / Q000378:metabolism; D000199:Actins / Q000235:genetics / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D009154:Mutation; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000821:virology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013696:Temperature; D017864:Tombusvirus / Q000235:genetics* / Q000502:physiology; D014779:Virus Replication
chemical_list: C102245:ARP3 protein, S cerevisiae; C465210:Act1 protein, S cerevisiae; D051378:Actin-Related Protein 3; D000199:Actins; D029701:Saccharomyces cerevisiae Proteins