Isolation and characterization of a split B-type DNA polymerase from the archaeon Methanobacterium thermoautotrophicum ΔH Journal Article
| Authors: | Kelman, Z.; Pietrokovski, S.; Hurwitz, J. |
| Article Title: | Isolation and characterization of a split B-type DNA polymerase from the archaeon Methanobacterium thermoautotrophicum ΔH |
| Abstract: | We describe here the isolation and characterization of a B-type DNA polymerase (PolB) from the archaeon Methanobacterium thermoautotrophicum ΔH. Uniquely, the catalytic domains of M. thermoautotrophicum PolB are encoded from two different genes, a feature that has not been observed as yet in other polymerases. The two genes were cloned, and the proteins were overexpressed in Escherichia coli and purified individually and as a complex. We demonstrate that both polypeptides are needed to form the active polymerase. Similar to other polymerases constituting the B-type family, PolB possesses both polymerase and 3'-5' exonuclease activities. We found that a homolog of replication protein A from M. thermoautotrophicum inhibits the PolB activity. The inhibition of DNA synthesis by replication protein A from M. thermoautotrophicum can be relieved by the addition of M. thermoautotrophicum homologs of replication factor C and proliferating cell nuclear antigen. The possible roles of PolB in M. thermoautotrophicum replication are discussed. |
| Keywords: | controlled study; dna-binding proteins; dna polymerase; dna replication; enzyme activity; bacteria (microorganisms); molecular cloning; cloning, molecular; bacterial protein; dna; amino acid sequence; molecular sequence data; enzyme analysis; nucleotide sequence; escherichia coli; recombinant proteins; base sequence; cycline; exonuclease; dna synthesis inhibition; archaea; replication protein a; electrophoresis, polyacrylamide gel; oligonucleotides; enzyme active site; dna polymerase beta; polypeptide; chromatography, affinity; single stranded dna binding protein; enzyme isolation; methanobacterium thermoautotrophicum; methanobacterium; priority journal; article; methanothermobacter thermautotrophicus |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 274 |
| Issue: | 40 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 1999-10-01 |
| Start Page: | 28751 |
| End Page: | 28761 |
| Language: | English |
| DOI: | 10.1074/jbc.274.40.28751 |
| PUBMED: | 10497247 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 16 August 2016 -- Source: Scopus |
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