Separation and partial characterization of two ribonucleic Acid polymerases from pea seedlings
D Glicklich1, J J Jendrisak, W M Becker
1Department of Botany and the Institute of Plant Development, University of Wisconsin, Madison, Wisconsin 53706.
Abstract:
Two DNA-dependent RNA polymerases (ribonucleoside triphosphate:RNA nucleotidyl transferase, EC 2.7.7.6) have been isolated from pea (Pisum sativum) seedlings. The enzymes were solubilized by sonication in high salt buffer and were separated by chromatography on diethylaminoethyl cellulose using a linear salt gradient. Polymerase I eluted at 0.10 m (NH(4))(2)SO(4), accounted for about 10% of the recovered activity and was completely insensitive to alpha-amanitin. Polymerase II eluted at 0.14 m (NH(4))(2)SO(4), accounted for the remaining 90% of recovered activity and was strongly inhibited by alpha-amanitin. Both enzymes preferred denatured to native DNA as template, both showed an absolute requirement of divalent cation, and both were sensitive to the ionic strength of the assay medium. The developing pea seedling seems a promising system for studies of possible changes in relative activities and roles of multiple RNA polymerases during eukaryotic development.
More Related Videos
06:31An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis
Published on: January 11, 2017
08:53Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
Published on: May 2, 2025
Related Concept Videos
Eukaryotic RNA Polymerases
All three eukaryotic RNAPs require specific transcription factors, of which the...
Restarting Stalled Replication Forks
Ribozymes
Ribozymes can be...
DNA Isolation
Lagging Strand Synthesis
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
