Mammalian Deoxyribonucleic Acid-dependent Ribonucleic Acid Polymerases
Journal of Biological ChemistryPublished 1 June 1973Open access
Bill Sugden, Walter Keller
Citations158
SJR quartileQ1
SJR score1.71
SNIP1.00
Generate an AI Snapshot to get a quick, structured summary of this paper.
Study Snapshot
ObjectiveStudy objective
MethodsResearch methodology
PopulationPopulation studied
Sample sizeSample sizes
OutcomesStudy outcomes here
ResultsStudy results comes here
LimitationsResearch study limitations comes here
A concise AI-generated summary of the paper will appear here once you click Generate AI Snapshot.
TL;DR
This work describes the partial purification of two stimulatory factors (SF-A and SF-B) which specifically stimulate the activity of RNA polymerase II in the presence of double-stranded DNA as template.
Abstract
Abstract A procedure is described for the purification of DNA-dependent RNA polymerase II of the human tissue culture cell lines HeLa and KB. The enzyme has a molecular weight of approximately 500,000 and in sodium dodecyl sulfate polyacrylamide gels invariably shows 3 subunits with molecular weights of 220,000, 140,000, and 35,000. In addition there perhaps is a fourth subunit of about 25,000 daltons or less in mass and a subunit of 170,000 daltons occurring in variable amounts. RNA polymerase II has a strong preference for single-stranded over double-stranded DNA as template and is most active in the presence of 1.5 to 3 mm Mn++ as divalent cation. We also describe the partial purification of two stimulatory factors (SF-A and SF-B) which specifically stimulate the activity of RNA polymerase II in the presence of double-stranded DNA as template.
Keywords
Biochemistry, Genetics and Molecular Biology
Journal of Biological ChemistryPROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENT
318,326 Citations1951OliverH. Lowry, NiraJ. Rosebrough +2 more
Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.
NatureCleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
251,640 Citations1970Ulrich K. Laemmli
Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Journal of Biological ChemistryThe Reliability of Molecular Weight Determinations by Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis
20,341 Citations1969K. Weber, Mary Osborn
The results show that the polyacrylamide gel electrophoresis method can be used with great confidence to determine the molecular weights of polypeptide chains for a wide variety of proteins.
ScienceAmino Acid Metabolism in Mammalian Cell Cultures
4,013 Citations1959Harry Eagle
The present article "is a progress report rather than a review and in large part summarizes studies from a single laboratory" on the minimal essential medium for cultivation of mammalian cells in either monolayer or suspension.
Journal of the American Chemical SocietyProcedures in Nucleic Acid Research.
2,042 Citations1967Waldo E. Cohn
NatureMultiple Forms of DNA-dependent RNA Polymerase in Eukaryotic Organisms
963 Citations1969Robert G. Roeder, William J. Rutter
Three distinct RNA polymerase activities have been isolated from developing sea urchin embryos, and in rat liver nuclei there are two RNA polymerases activities.
Journal of Biological ChemistryA New Method for the Large Scale Purification of Escherichia coli Deoxyribonucleic Acid-dependent Ribonucleic Acid Polymerase
955 Citations1969Richard R. Burgess
A method for the purification of Escherichia coli DNA-dependent RNA polymerase which is rapid, reproducible, high in yield, and able to handle preparations using from 1 g to 3 kg of cells is described.
ScienceSpecific Inhibition of Nuclear RNA Polymerase II by α-Amanitin
868 Citations1970Thomas J. Lindell, Fanyela Weinberg +3 more
α-Amanitin, a toxic substance from the mushroom Amanita phalloides, is a potent inhibitor of DNA-dependent RNA polymerase II (the nucleoplasmic form) from sea urchin, rat liver, and calf thymus.
Methods in enzymology on CD-ROM/Methods in enzymology[40] Thin-layer separation methods for nucleic acid derivatives
505 Citations1967Kurt Randerath, Erika Randerath
Thin-layer separation methods for nucleic acid derivatives that are found effective in laboratory are described, including PEI-cellulose anion-exchange layers, which afford particularly sharp and highly reproducible separations of nucleotides.
Biochemical and Biophysical Research Communicationsα-Amanitin: A specific inhibitor of one of two DNA-dependent RNA polymerase activities from calf thymus
488 Citations1970C. Kédinger, Marek Gniazdowski +3 more
Experimental Biology and MedicinePropagation in a Fluid Medium of a Human Epidermoid Carcinoma, Strain KB
482 Citations1955Harry Eagle
An epidermoid carcinoma of the floor of the mouth was cultivated directly onto a glass surface in a medium consisting of 13 amino acids, 7 vitamins, glucose, salts, and 10% human serum, and all were used at concentrations permitting maximal growth of that cell.
Proceedings of the National Academy of SciencesSpecific Nucleolar and Nucleoplasmic RNA Polymerases
451 Citations1970Robert G. Roeder, William J. Rutter
It is concluded that the RNA polymerases are specifically localized within the nucleus and may, therefore, play specific roles in the regulation of genetic transcription.
Proceedings of the National Academy of SciencesTHE CYCLIC HELIX AND CYCLIC COIL FORMS OF POLYOMA VIRAL DNA
294 Citations1963Roger Weil, Jerome Vinograd
Evidence is given suggesting that a fraction of the polyoma DNA molecules are cyclic, which would in part account for the properties of the DNA extracted from polyoma virus.
Journal of Molecular BiologyAmount of viral DNA in the genome of cells transformed by adenovirus type 2
292 Citations1973Ulf Pettersson, Joe Sambrook
The number of copies of viralDNA in DNA of cells transformed by adenovirus type 2 has been determined by following the kinetics of reassociation of 32P-labeled viral DNA in the presence of unlabeled DNA extracted from transformed and control cells.
ScienceEnzyme from Calf Thymus Degrading the RNA Moiety of DNA-RNA Hybrids: Effect on DNA-Dependent RNA Polymerase
261 Citations1969Hans Stein, Peter Hausen
If prepared free of the hybrid-degrading enzyme, RNA polymerase from calf thymus shows a fivefold increase in activity on denatured DNA as compared to native DNA.
Nature New BiologyDNA Polymerases from Human Cells
257 Citations1971Arthur Weissbach, A J Schlabach +2 more
The same distribution of DNA polymerases has been found in the nucleus and cytoplasm of WI-38 cells, a normal diploid human cell line.
Journal of Molecular BiologyRibosomal RNA synthesis in isolated nuclei
245 Citations1972Ronald H. Reeder, Robert G. Roeder
It is suggested that ribosomal RNA is synthesized by form I, the major α-amanitin-resistant RNA polymerase found in these nuclei, but cannot rule out the possible involvement of form III, a minor α-Manatin-resistantRNA polymerase which is also present.
Proceedings of the National Academy of SciencesDegradation of DNA RNA Hybrids by Ribonuclease H and DNA Polymerases of Cellular and Viral Origin
222 Citations1972Walter Keller, Robert J. Crouch
Ribonuclease H from human KB cells, chick embryos, calf thymus, avian myeloblastosis virus, and Rous associated virus specifically degrades the RNA of DNA, which is an intrinsic part of the viral DNA polymerase.
Proceedings of the National Academy of SciencesProducts of RNA Polymerases in HeLa Cell Nuclei
221 Citations1971Ester A. Zylber, Sheldon Penman
Journal of Biological ChemistryLow Molecular Weight Deoxyribonucleic Acid Polymerase in Mammalian Cells
189 Citations1971Lucy M.S. Chang, F.J. Bollum
The 3.3 S enzyme extracted from rabbit bone marrow uses template information for nucleotide selection and is therefore a DNA polymerase and not a terminal deoxynucleotidyltransferase, which differs from the 6 to 8 SDNA polymerase in its behavior on phosphocellulose and its optimum activity at alkaline pH.
European Journal of BiochemistryAnimal DNA‐Dependent RNA Polymerases
153 Citations1972Madeleine Mellhac, Z Tyśper +1 more
European Journal of BiochemistryAnimal DNA‐Dependent RNA Polymerases
137 Citations1972Claude Kédinger, Pierre Chambon
Journal of VirologyTranscription of the Adenovirus Genome by an α-Amanitine-Sensitive Ribonucleic Acid Polymerase in HeLa Cells
134 Citations1972Richard P. Price, Sheldon Penman
The properties of the ribonucleic acid (RNA) polymerase activity which transcribes the major portion of the adenovirus genome were studied and its product is high-molecular-weight heterogeneous RNA.
BiochemistryPhysical studies on ribonucleic acid polymerase from Escherichia coli B
123 Citations1970Michael J. Chamberlin, Darwin K. Berg
Proceedings of the National Academy of SciencesMolecular Structures of DNA-Dependent RNA Polymerases (II) from Calf Thymus and Rat Liver
112 Citations1971Robert F. Weaver, S. P. Blatti +1 more
The general structural similarity of the eukaryotic RNA polymerase II with that of the prokaryotic polymerase suggests that the modes of action and regulation may be analogous, until unequivocal evidence is obtained on this point.
Journal of VirologyState of Adenovirus 2 Deoxyribonucleic Acid in the Nucleus and Its Mode of Transcription: Studies with Isolated Viral Deoxyribonucleic Acid-Protein Complexes and Isolated Nuclei
99 Citations1972Robert D. Wallace, Joseph R. Kates
Newly replicated adenovirus 2 deoxyribonucleic acid (DNA) can be isolated from the nucleus of HeLa cells by a gentle lysis procedure as a fairly homogeneous complex with a sedimentation of 73S and the synthesis of virus-specific RNA by the isolated nuclei is strongly inhibited by low doses of alpha-amanitine.
ScienceEncapsidation of Free Host DNA by Simian Virus 40: A Simian Virus 40 Pseudovirus
67 Citations1970David Trilling, David Axelrod
Under specified growth conditions, simian virus 40 encapsidated host DNA in a noncircular form free of viral DNA under cesium chloride equilibrium density centrifugation.
Journal of VirologyInhibition by α-Amanitin of Simian Virus 40-Specific Ribonucleic Acid Synthesis in Nuclei of Infected Monkey Cells
57 Citations1972Arlene Hildebrandt Jackson, Bill Sugden
Results show that an alpha-amanitin-sensitive RNA polymerase activity is responsible for transcription of simian virus 40 deoxyribonucleic acid.
VirologyStructural proteins of adenoviruses
56 Citations1968Ulf Pettersson, Lennart Philipson +1 more
Cold Spring Harbor Symposia on Quantitative BiologyA Factor Stimulating the Transcription on Double-Stranded DNA by Purified RNA Polymerase from Rat Liver Nuclei
42 Citations1970Klaus H. Seifart
Cold Spring Harbor Symposia on Quantitative BiologyFactors Influencing the Activity of Mammalia RNA Polymerase
40 Citations1970Hans Stein, Peter Hausen
Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein SynthesisRNA polymerase of HeLa cells
26 Citations1967Akira Ishihama
A simple procedure to obtain DNA-free RNA polymerase (nucleosidetriphosphate: RNA nucleotidyltransferase, EC 2.7.6) from HeLa cells was developed from the preparation of extracts in a hypotonic medium in a French pressure cell, exhibiting similar reaction profiles to those of Escherichia coli RNA polymerases.
Cold Spring Harbor Symposia on Quantitative BiologyMammalian RNA Polymerase: Structural and Functional Properties
23 Citations1970Walter Keller, Ronald S. Goor
Cold Spring Harbor Symposia on Quantitative BiologyStudies on DNA-Directed RNA Polymerase; Formation of DNA-RNA Complexes with Single-Stranded X 174 DNA as Template
20 Citations1963Michael J. Chamberlin, Paul Berg
Cold Spring Harbor Symposia on Quantitative BiologyRNA Polymerase from HeLa Cells
17 Citations1970Bill Sugden, Joe Sambrook