The use of fungal in vitro systems for studying translational regulation.
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.
Wu
Cheng
C
Department of Environmental and Biomolecular Systems, OGI School of Science and Engineering, Oregon Health and Science University, Beaverton, Oregon, USA.
Amrani
Nadia
N
Jacobson
Allan
A
Sachs
Matthew S
MS
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
D017931:DNA Primers; D010454:Peptide Termination Factors; D029701:Saccharomyces cerevisiae Proteins; C087971:peptide-chain-release factor 3; D003513:Cycloheximide; D008836:Micrococcal Nuclease