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Subunits of the Schizosaccharomyces pombe RNA polymerase II: enzyme purification and structure of the subunit 3 gene.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Azuma, Yoshiaki Yamagishi, Masahiro Ishihama, Akira |
| Copyright Year | 1993 |
| Abstract | To improve our understanding of the structure and function of eukaryotic RNA polymerase II, we purified the enzyme from the fission yeast Schizosaccharomyces pombe. The highly purified RNA polymerase II contained more than eleven polypeptides. The sizes of the largest the second-, and the third-largest polypeptides as measured by SDS-polyacrylamide gel electrophoresis were about 210, 150, and 40 kilodaltons (kDa), respectively, and are similar to those of RPB1, 2, and 3 subunits of Saccharomyces cerevisiae RNA polymerase II. Using the degenerated primers designed after amino acid micro-sequencing of the 40 kDa third-largest polypeptide (subunit 3), we cloned the subunit 3 gene (rpb3) and determined its DNA sequence. Taken together with the sequence of parts of PCR-amplified cDNA, the predicted coding sequence of rpb3, interrupted by two introns, was found to encode a polypeptide of 297 amino acid residues in length with a molecular weight of 34 kDa. The S. pombe subunit 3 contains four structural domains conserved for the alpha-subunit family of RNA polymerase from both eukaryotes and prokaryotes. A putative leucine zipper motif was found to exist in the C-terminal proximal conserved region (domain D). Possible functions of the conserved domains are discussed. |
| Starting Page | 1 |
| Ending Page | 2 |
| Page Count | 2 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://oup.silverchair-cdn.com/oup/backfile/Content_public/Journal/nar/21/16/10.1093_nar_21.16.3749/2/21-16-3749.pdf?Expires=1496220559&Key-Pair-Id=APKAIUCZBIA4LVPAVW3Q&Signature=POAOQzj4-bU6IHn4LyVUE~SY6fjp8n8XtztJZGyLWsKnrm0A3jrDLcTqCmGGgoYlsAVdpElNqEmV1CfZqtGwjDrXvNMXoSSwbah2hXWvI4w2~JBF2tW2U9-azlMU~RvCnIAt26ILOCVqrVMnPo7PM0IH7yaRSa~rhgClNxvjEmyUc7tCm7lOAndYFWjy8bE6Fkgg07m6AhHAUtDCTmGH3tCH0Qa4GomGnDZtCVy1D7aYFLOBvrO0tOpZxLk3K-KPcjeo2YWRVXQLDLDqPdqMs9Y-PNDSXpoVoaEZBeNhcP4gj6Cu49pCR1RtSSW2TI3Oi~UAkgL9cTRD~197ZJDb5Q__ |
| PubMed reference number | 8367291v1 |
| Volume Number | 21 |
| Issue Number | 16 |
| Journal | Nucleic acids research |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Amino Acids Biopolymer Sequencing Clone Cells Contain (action) DNA, Complementary DNA-Directed RNA Polymerase Encode (action) Gel Electrophoresis (lab technique) Introns Largest Leucine Zippers Molecular Weight Open Reading Frames Polyacrylamide Gel Electrophoresis Polymerase Chain Reaction Polypeptides Prokaryote Protein Domain RNA Polymerase II Schizosaccharomyces pombe polyacrylamide gels |
| Content Type | Text |
| Resource Type | Article |
