EurekaMag.com logo
+ Site Statistics
References:
53,869,633
Abstracts:
29,686,251
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Biochemical analysis of replication factor C from the hyperthermophilic archaeon Pyrococcus furiosus



Biochemical analysis of replication factor C from the hyperthermophilic archaeon Pyrococcus furiosus



Journal of Bacteriology 183(8): 2614-2623



Replication factor C (RFC) and proliferating cell nuclear antigen (PCNA) are accessory proteins essential for processive DNA synthesis in the domain Eucarya. The function of RFC is to load PCNA, a processivity factor of eukaryotic DNA polymerases delta and epsilon, onto primed DNA templates. RFC-like genes, arranged in tandem in the Pyrococcus furiosus genome, were cloned and expressed individually in Escherichia coli cells to determine their roles in DNA synthesis. The P. furiosus RFC (PfuRFC) consists of a small subunit (RFCS) and a large subunit (RFCL). Highly purified RFCS possesses an ATPase activity, which was stimulated up to twofold in the presence of both single-stranded DNA (ssDNA) and P. furiosus PCNA (PfuPCNA). The ATPase activity of PfuRFC itself was as strong as that of RFCS. However, in the presence of PfuPCNA and ssDNA, PfuRFC exhibited a 10-fold increase in ATPase activity under the same conditions. RFCL formed very large complexes by itself and had an extremely weak ATPase activity, which was not stimulated by PfuPCNA and DNA. The PfuRFC stimulated PfuPCNA-dependent DNA synthesis by both polymerase I and polymerase II from P. furiosus. We propose that PfuRFC is required for efficient loading of PfuPCNA and that the role of RFC in processive DNA synthesis is conserved in Archaea and Eucarya.

(PDF emailed within 0-6 h: $19.90)

Accession: 009778701

Download citation: RISBibTeXText

PMID: 11274122

DOI: 10.1128/JB.183.8.2614-2623.2001



Related references

Transcriptional and biochemical analysis of starch metabolism in the hyperthermophilic archaeon Pyrococcus furiosus. Journal of Bacteriology 188(6): 2115-2125, 2006

Biochemical characterization of endonuclease V from the hyperthermophilic archaeon, Pyrococcus furiosus. Journal of Biochemistry 155(5): 325-333, 2015

Biochemical adaptations of two sugar kinases from the hyperthermophilic archaeon Pyrococcus furiosus. Biochemical Journal 366(Pt 1): 121-127, 2002

Molecular and biochemical characterization of the ADP-dependent phosphofructokinase from the hyperthermophilic archaeon Pyrococcus furiosus. Journal of Biological Chemistry 274(30): 21023-8, 1999

Molecular and biochemical characterization of an endo-b-1,3-glucanase of the hyperthermophilic archaeon Pyrococcus furiosus. The Journal of Biological Chemistry 272: 258-64, 1997

Localized melting of duplex DNA by Cdc6/Orc1 at the DNA replication origin in the hyperthermophilic archaeon Pyrococcus furiosus. Extremophiles 14(1): 21-31, 2010

Molecular and biochemical characterization of an endo-beta-1,3- glucanase of the hyperthermophilic archaeon Pyrococcus furiosus. Journal of Biological Chemistry 272(50): 31258-31264, 1998

Biochemical evidence for the presence of two alpha-glucoside ABC-transport systems in the hyperthermophilic archaeon Pyrococcus furiosus. Archaea 1(1): 19-25, 2005

Genetic and biochemical characterization of a short-chain alcohol dehydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus. European Journal of Biochemistry 268(10): 3062-3068, 2001

Experimental and computational analysis of the secretome of the hyperthermophilic archaeon Pyrococcus furiosus. Extremophiles 17(6): 921-930, 2014

Purification and sequence analysis of sulfhydrogenase II from the hyperthermophilic archaeon, Pyrococcus furiosus. Abstracts of the General Meeting of the American Society for Microbiology 99: 421, 1999

Defining components of the chromosomal origin of replication of the hyperthermophilic archaeon Pyrococcus furiosus needed for construction of a stable replicating shuttle vector. Applied and Environmental Microbiology 77(18): 6343-6349, 2011

Microarray analysis of the hyperthermophilic archaeon Pyrococcus furiosus exposed to gamma irradiation. Extremophiles 11(1): 19-29, 2006

Reverse gyrase from the hyperthermophilic archaeon, Pyrococcus furiosus Cloning, sequencing and transcript analysis. Abstracts of the General Meeting of the American Society for Microbiology 96(0): 303, 1996