Biomaterial Database

Yeast DNA-dependent RNA polymerase I. A rapid procedure for the large scale purification of homogeneous enzyme. 1976

P Valenzuela, and F Weinberg, and G Bell, and W J Rutter

A procedure has been developed for the rapid purification of large amounts of yeast RNA polymerase I (A). The method involves batchwise treatment with phosphocellulose and DEAE-cellulose, ion filtration chromatography on DEAE-Sephadex, sucrose gradient centrifugation, and DNA-cellulose chromatography. The enzyme obtained is apparently homogeneous by sedimentation velocity analysis and has a specific activity of 300 nmol of UMP incorporated into RNA in 10 min per mg of protein. Between 30 and 45 mg of enzyme can be obtained in 5 days from 3.0 kg of yeast cells. The subunit composition of the enzyme was determined by polyacrylamide gel electrophoresis in the presence of 0.1% sodium dodecyl sulfate. The purified polymerase is composed of 11 putative subunits with molecular weights 185,000 (Ia), 137,000 (Ib), 48,000 (Ic), 44,000 (Id), 41,000 (Ie), 36,000 (If), 28,000 (Ig), 24,000 (Ih), 20,000 (Ii), 14,500 (Ij), and 12,000 (Ik). Yeast polymerase I separates into two forms when subjected to gel electrophoresis under nondenaturing conditions. The main component which migrates faster contains all the subunits except the polypeptides Ic and If. The slow migrating component which is present in lower amounts contains all the subunits.

UI	MeSH Term	Description	Entries
D008970	Molecular Weight	The sum of the weight of all the atoms in a molecule.	Molecular Weights,Weight, Molecular,Weights, Molecular
D004591	Electrophoresis, Polyacrylamide Gel	Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.	Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D012321	DNA-Directed RNA Polymerases	Enzymes that catalyze DNA template-directed extension of the 3'-end of an RNA strand one nucleotide at a time. They can initiate a chain de novo. In eukaryotes, three forms of the enzyme have been distinguished on the basis of sensitivity to alpha-amanitin, and the type of RNA synthesized. (From Enzyme Nomenclature, 1992).	DNA-Dependent RNA Polymerases,RNA Polymerases,Transcriptases,DNA-Directed RNA Polymerase,RNA Polymerase,Transcriptase,DNA Dependent RNA Polymerases,DNA Directed RNA Polymerase,DNA Directed RNA Polymerases,Polymerase, DNA-Directed RNA,Polymerase, RNA,Polymerases, DNA-Dependent RNA,Polymerases, DNA-Directed RNA,Polymerases, RNA,RNA Polymerase, DNA-Directed,RNA Polymerases, DNA-Dependent,RNA Polymerases, DNA-Directed
D012441	Saccharomyces cerevisiae	A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.	Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D046911	Macromolecular Substances	Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.	Macromolecular Complexes,Macromolecular Compounds,Macromolecular Compounds and Complexes,Complexes, Macromolecular,Compounds, Macromolecular,Substances, Macromolecular