Cloning of the large subunit of replication protein A (RPA) from yeast Saccharomyces cerevisiae and its DNA binding activity through redox potential

Haeng Soon Jeong, In Chel Jeong, Andre Kim, Shin Won Kang, Sung Kang Ho, Yung Jin Kim, Suk-Hee Lee, Jang Su Park

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Eukaryotic replication protein A (RPA) is a single-stranded( ss) DNA binding protein with multiple functions in DNA replication, repair, and genetic recombination. The 70-kDa subunit of eukaryotic RPA contains a conserved four cysteine-type zinc-finger motif that has been implicated in the regulation of DNA replication and repair. Recently, we described a novel function for the zinc-finger motif in the regulation of human RPA's ssDNA binding activity through reduction-oxidation (redox). Here, we show that yeast RPA's ssDNA binding activity is regulated by redox potential through its RPA32 and/or RPA14 subunits. Yeast RPA requires a reducing agent, such as dithiothreitol (DTT), for its ssDNA binding activity. Also, under non-reducing conditions, its DNA binding activity decreases 20 fold. In contrast, the RPA70 subunit does not require DTT for its DNA binding activity and is not affected by the redox condition. These results suggest that all three subunits are required for the regulation of RPA's DNA binding activity through redox potential.

Original languageEnglish
Pages (from-to)194-198
Number of pages5
JournalJournal of Biochemistry and Molecular Biology
Volume35
Issue number2
StatePublished - Mar 2002

Fingerprint

Replication Protein A
Cloning
Yeast
Saccharomyces cerevisiae
Organism Cloning
Thermodynamic properties
Yeasts
Dithiothreitol
Zinc Fingers
DNA Replication
DNA Repair
Oxidation
DNA
Fungal Proteins
Reducing Agents
DNA-Binding Proteins
Zinc
Repair
Genetic Recombination
Cysteine

Keywords

  • Redox
  • Replication protein A (RPA)
  • RPA70
  • Single-stranded DNA binding protein

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Cloning of the large subunit of replication protein A (RPA) from yeast Saccharomyces cerevisiae and its DNA binding activity through redox potential. / Jeong, Haeng Soon; Jeong, In Chel; Kim, Andre; Kang, Shin Won; Ho, Sung Kang; Kim, Yung Jin; Lee, Suk-Hee; Park, Jang Su.

In: Journal of Biochemistry and Molecular Biology, Vol. 35, No. 2, 03.2002, p. 194-198.

Research output: Contribution to journalArticle

Jeong, Haeng Soon ; Jeong, In Chel ; Kim, Andre ; Kang, Shin Won ; Ho, Sung Kang ; Kim, Yung Jin ; Lee, Suk-Hee ; Park, Jang Su. / Cloning of the large subunit of replication protein A (RPA) from yeast Saccharomyces cerevisiae and its DNA binding activity through redox potential. In: Journal of Biochemistry and Molecular Biology. 2002 ; Vol. 35, No. 2. pp. 194-198.
@article{8a42682afb7242e4b62e3dbca5e1e498,
title = "Cloning of the large subunit of replication protein A (RPA) from yeast Saccharomyces cerevisiae and its DNA binding activity through redox potential",
abstract = "Eukaryotic replication protein A (RPA) is a single-stranded( ss) DNA binding protein with multiple functions in DNA replication, repair, and genetic recombination. The 70-kDa subunit of eukaryotic RPA contains a conserved four cysteine-type zinc-finger motif that has been implicated in the regulation of DNA replication and repair. Recently, we described a novel function for the zinc-finger motif in the regulation of human RPA's ssDNA binding activity through reduction-oxidation (redox). Here, we show that yeast RPA's ssDNA binding activity is regulated by redox potential through its RPA32 and/or RPA14 subunits. Yeast RPA requires a reducing agent, such as dithiothreitol (DTT), for its ssDNA binding activity. Also, under non-reducing conditions, its DNA binding activity decreases 20 fold. In contrast, the RPA70 subunit does not require DTT for its DNA binding activity and is not affected by the redox condition. These results suggest that all three subunits are required for the regulation of RPA's DNA binding activity through redox potential.",
keywords = "Redox, Replication protein A (RPA), RPA70, Single-stranded DNA binding protein",
author = "Jeong, {Haeng Soon} and Jeong, {In Chel} and Andre Kim and Kang, {Shin Won} and Ho, {Sung Kang} and Kim, {Yung Jin} and Suk-Hee Lee and Park, {Jang Su}",
year = "2002",
month = "3",
language = "English",
volume = "35",
pages = "194--198",
journal = "BMB Reports",
issn = "1976-6696",
publisher = "The Biochemical Society of the Republic of Korea",
number = "2",

}

TY - JOUR

T1 - Cloning of the large subunit of replication protein A (RPA) from yeast Saccharomyces cerevisiae and its DNA binding activity through redox potential

AU - Jeong, Haeng Soon

AU - Jeong, In Chel

AU - Kim, Andre

AU - Kang, Shin Won

AU - Ho, Sung Kang

AU - Kim, Yung Jin

AU - Lee, Suk-Hee

AU - Park, Jang Su

PY - 2002/3

Y1 - 2002/3

N2 - Eukaryotic replication protein A (RPA) is a single-stranded( ss) DNA binding protein with multiple functions in DNA replication, repair, and genetic recombination. The 70-kDa subunit of eukaryotic RPA contains a conserved four cysteine-type zinc-finger motif that has been implicated in the regulation of DNA replication and repair. Recently, we described a novel function for the zinc-finger motif in the regulation of human RPA's ssDNA binding activity through reduction-oxidation (redox). Here, we show that yeast RPA's ssDNA binding activity is regulated by redox potential through its RPA32 and/or RPA14 subunits. Yeast RPA requires a reducing agent, such as dithiothreitol (DTT), for its ssDNA binding activity. Also, under non-reducing conditions, its DNA binding activity decreases 20 fold. In contrast, the RPA70 subunit does not require DTT for its DNA binding activity and is not affected by the redox condition. These results suggest that all three subunits are required for the regulation of RPA's DNA binding activity through redox potential.

AB - Eukaryotic replication protein A (RPA) is a single-stranded( ss) DNA binding protein with multiple functions in DNA replication, repair, and genetic recombination. The 70-kDa subunit of eukaryotic RPA contains a conserved four cysteine-type zinc-finger motif that has been implicated in the regulation of DNA replication and repair. Recently, we described a novel function for the zinc-finger motif in the regulation of human RPA's ssDNA binding activity through reduction-oxidation (redox). Here, we show that yeast RPA's ssDNA binding activity is regulated by redox potential through its RPA32 and/or RPA14 subunits. Yeast RPA requires a reducing agent, such as dithiothreitol (DTT), for its ssDNA binding activity. Also, under non-reducing conditions, its DNA binding activity decreases 20 fold. In contrast, the RPA70 subunit does not require DTT for its DNA binding activity and is not affected by the redox condition. These results suggest that all three subunits are required for the regulation of RPA's DNA binding activity through redox potential.

KW - Redox

KW - Replication protein A (RPA)

KW - RPA70

KW - Single-stranded DNA binding protein

UR - http://www.scopus.com/inward/record.url?scp=0036004553&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036004553&partnerID=8YFLogxK

M3 - Article

VL - 35

SP - 194

EP - 198

JO - BMB Reports

JF - BMB Reports

SN - 1976-6696

IS - 2

ER -