title: The use of fungal in vitro systems for studying translational regulation.
abstract: The use of cell-free systems enables biochemical determination of factors and mechanisms contributing to translational processes. The preparation and use of cell-free translation systems from the fungi Saccharomyces cerevisiae and Neurospora crassa are described. Examples provided illustrate the use of these systems, in conjunction with luciferase assays, [(35)S]Met incorporation, and primer-extension inhibition (toeprint) analyses, to assess the translational effects of upstream open reading frames and premature termination codons.
authors.affiliation: Department of Environmental and Biomolecular Systems, OGI School of Science and Engineering, Oregon Health and Science University, Beaverton, Oregon, USA.
authors.forename: Cheng
authors.initials: C
authors.lastname: Wu
authors.forename: Nadia
authors.initials: N
authors.lastname: Amrani
authors.forename: Allan
authors.initials: A
authors.lastname: Jacobson
authors.forename: Matthew S
authors.initials: MS
authors.lastname: Sachs
mesh_terms: D002474:Cell-Free System; D003513:Cycloheximide / Q000494:pharmacology; D017931:DNA Primers; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D008836:Micrococcal Nuclease; D009492:Neurospora crassa / Q000502:physiology*; D016366:Open Reading Frames / Q000502:physiology; D010443:Peptide Chain Termination, Translational; D010454:Peptide Termination Factors / Q000502:physiology; D014176:Protein Biosynthesis / Q000502:physiology; D012270:Ribosomes / Q000187:drug effects; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology
chemical_list: D017931:DNA Primers; D010454:Peptide Termination Factors; D029701:Saccharomyces cerevisiae Proteins; C087971:peptide-chain-release factor 3; D003513:Cycloheximide; D008836:Micrococcal Nuclease