Identification of the critical region in Replication factor C from Pyrococcus furiosus for the stable complex formation with Proliferating cell nuclear antigen and DNA

Hirokazu Nishida, Sonoko Ishino, Tomoko Miyata, Kosuke Morikawa, Yoshizumi Ishino

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Replication factor C (RFC) and proliferating cell nuclear antigen (PCNA) are accessory proteins essential for processive DNA synthesis. The function of RFC is to load PCNA, a processivity factor of replicative DNA polymerases, onto primed DNA templates. The central hole of the PCNA homo-trimeric ring encircles doublestranded DNA, so that DNA polymerases can operate for DNA synthesis with PCNA along a DNA template. The Pyrococcus furiosus RFC (PfuRFC) consists of a small subunit (RFCS, 37kDa) and a large subunit (RFCL, 55kDa), which show significant sequence identity to the eukaryotic homologs. The C-terminal region of RFCL has an acidic cluster of about 30 amino acids, which consists mainly of glutamic acid residues, and a following basic cluster of 10 amino acids, which consists mainly of lysine residues. These clusters of charged amino acids, which precede the C-terminal consensus sequence, PIP (PCNA interacting protein)-box, are conserved in several archaeal RFCLs. The series of mutant PfuRFC containing the C-terminal deletions in RFCL were constructed. The mutational analyses showed that the charged cluster is not essential for loading of PCNA onto DNA. However, the region containing the basic cluster is important for the stable ternary (RFC-PCNA-DNA) complex formation.

Original languageEnglish
Pages (from-to)83-93
Number of pages11
JournalGenes and Genetic Systems
Volume80
Issue number2
DOIs
Publication statusPublished - Oct 3 2005

Fingerprint

Pyrococcus furiosus
Replication Protein C
proliferating cell nuclear antigen
Proliferating Cell Nuclear Antigen
nuclear genome
DNA
DNA-directed DNA polymerase
DNA-Directed DNA Polymerase
Amino Acids
amino acids
synthesis
consensus sequence
Consensus Sequence
Accessories
Homo
glutamic acid
Lysine
Glutamic Acid
lysine
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

@article{b91a7bc390494013a49d88f99f5c0145,
title = "Identification of the critical region in Replication factor C from Pyrococcus furiosus for the stable complex formation with Proliferating cell nuclear antigen and DNA",
abstract = "Replication factor C (RFC) and proliferating cell nuclear antigen (PCNA) are accessory proteins essential for processive DNA synthesis. The function of RFC is to load PCNA, a processivity factor of replicative DNA polymerases, onto primed DNA templates. The central hole of the PCNA homo-trimeric ring encircles doublestranded DNA, so that DNA polymerases can operate for DNA synthesis with PCNA along a DNA template. The Pyrococcus furiosus RFC (PfuRFC) consists of a small subunit (RFCS, 37kDa) and a large subunit (RFCL, 55kDa), which show significant sequence identity to the eukaryotic homologs. The C-terminal region of RFCL has an acidic cluster of about 30 amino acids, which consists mainly of glutamic acid residues, and a following basic cluster of 10 amino acids, which consists mainly of lysine residues. These clusters of charged amino acids, which precede the C-terminal consensus sequence, PIP (PCNA interacting protein)-box, are conserved in several archaeal RFCLs. The series of mutant PfuRFC containing the C-terminal deletions in RFCL were constructed. The mutational analyses showed that the charged cluster is not essential for loading of PCNA onto DNA. However, the region containing the basic cluster is important for the stable ternary (RFC-PCNA-DNA) complex formation.",
author = "Hirokazu Nishida and Sonoko Ishino and Tomoko Miyata and Kosuke Morikawa and Yoshizumi Ishino",
year = "2005",
month = "10",
day = "3",
doi = "10.1266/ggs.80.83",
language = "English",
volume = "80",
pages = "83--93",
journal = "Genes and Genetic Systems",
issn = "1341-7568",
publisher = "The Genetics Society of Japan",
number = "2",

}

TY - JOUR

T1 - Identification of the critical region in Replication factor C from Pyrococcus furiosus for the stable complex formation with Proliferating cell nuclear antigen and DNA

AU - Nishida, Hirokazu

AU - Ishino, Sonoko

AU - Miyata, Tomoko

AU - Morikawa, Kosuke

AU - Ishino, Yoshizumi

PY - 2005/10/3

Y1 - 2005/10/3

N2 - Replication factor C (RFC) and proliferating cell nuclear antigen (PCNA) are accessory proteins essential for processive DNA synthesis. The function of RFC is to load PCNA, a processivity factor of replicative DNA polymerases, onto primed DNA templates. The central hole of the PCNA homo-trimeric ring encircles doublestranded DNA, so that DNA polymerases can operate for DNA synthesis with PCNA along a DNA template. The Pyrococcus furiosus RFC (PfuRFC) consists of a small subunit (RFCS, 37kDa) and a large subunit (RFCL, 55kDa), which show significant sequence identity to the eukaryotic homologs. The C-terminal region of RFCL has an acidic cluster of about 30 amino acids, which consists mainly of glutamic acid residues, and a following basic cluster of 10 amino acids, which consists mainly of lysine residues. These clusters of charged amino acids, which precede the C-terminal consensus sequence, PIP (PCNA interacting protein)-box, are conserved in several archaeal RFCLs. The series of mutant PfuRFC containing the C-terminal deletions in RFCL were constructed. The mutational analyses showed that the charged cluster is not essential for loading of PCNA onto DNA. However, the region containing the basic cluster is important for the stable ternary (RFC-PCNA-DNA) complex formation.

AB - Replication factor C (RFC) and proliferating cell nuclear antigen (PCNA) are accessory proteins essential for processive DNA synthesis. The function of RFC is to load PCNA, a processivity factor of replicative DNA polymerases, onto primed DNA templates. The central hole of the PCNA homo-trimeric ring encircles doublestranded DNA, so that DNA polymerases can operate for DNA synthesis with PCNA along a DNA template. The Pyrococcus furiosus RFC (PfuRFC) consists of a small subunit (RFCS, 37kDa) and a large subunit (RFCL, 55kDa), which show significant sequence identity to the eukaryotic homologs. The C-terminal region of RFCL has an acidic cluster of about 30 amino acids, which consists mainly of glutamic acid residues, and a following basic cluster of 10 amino acids, which consists mainly of lysine residues. These clusters of charged amino acids, which precede the C-terminal consensus sequence, PIP (PCNA interacting protein)-box, are conserved in several archaeal RFCLs. The series of mutant PfuRFC containing the C-terminal deletions in RFCL were constructed. The mutational analyses showed that the charged cluster is not essential for loading of PCNA onto DNA. However, the region containing the basic cluster is important for the stable ternary (RFC-PCNA-DNA) complex formation.

UR - http://www.scopus.com/inward/record.url?scp=25144451780&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=25144451780&partnerID=8YFLogxK

U2 - 10.1266/ggs.80.83

DO - 10.1266/ggs.80.83

M3 - Article

C2 - 16172520

AN - SCOPUS:25144451780

VL - 80

SP - 83

EP - 93

JO - Genes and Genetic Systems

JF - Genes and Genetic Systems

SN - 1341-7568

IS - 2

ER -