title: Specific initiation by RNA polymerase I in a whole-cell extract from yeast.
abstract: A protocol is described for making a soluble whole-cell extract from yeast (Saccharomyces cerevisiae) that supports active and specific transcription initiation by RNA polymerases I, II, and III. Specific initiation by polymerase I decreases in high-density cultures, paralleling the decrease in abundance of the endogenous 35S rRNA precursor. This extract should be useful for studying the molecular mechanisms that regulate rRNA transcription in yeast.
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
authors.forename: M C
authors.initials: MC
authors.lastname: Schultz
authors.forename: S Y
authors.initials: SY
authors.lastname: Choe
authors.forename: R H
authors.initials: RH
authors.lastname: Reeder
mesh_terms: D001483:Base Sequence; D003600:Cytosol / Q000378:metabolism; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D012318:RNA Polymerase I / Q000378:metabolism*; D012322:RNA Precursors / Q000378:metabolism; D012335:RNA, Ribosomal / Q000235:genetics*; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D015345:Oligonucleotide Probes; D012322:RNA Precursors; D012335:RNA, Ribosomal; D012318:RNA Polymerase I