34 results match your criteria rpa14 subunits


Generation of Fluorescent Versions of Saccharomyces cerevisiae RPA to Study the Conformational Dynamics of Its ssDNA-Binding Domains.

Methods Mol Biol 2021 ;2281:151-168

Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, MO, USA.

Replication protein A (RPA) is an essential single-stranded DNA (ssDNA)-binding protein that sequesters ssDNA and protects it from nucleolytic degradation. The RPA-ssDNA nucleoprotein acts as a hub to recruit over two dozen DNA metabolic enzymes onto ssDNA to coordinate DNA replication, repair, and recombination. RPA functions as a heterotrimer composed of RPA70, RPA32, and RPA14 subunits and has multiple DNA-binding and protein-interaction domains. Read More

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January 2021

Determinants of replication protein A subunit interactions revealed using a phosphomimetic peptide.

J Biol Chem 2020 12 30;295(52):18449-18458. Epub 2020 Oct 30.

Department of Chemistry, Gwangju Institute of Science and Technology, Gwangju, Republic of Korea. Electronic address:

Replication protein A (RPA) is a eukaryotic ssDNA-binding protein and contains three subunits: RPA70, RPA32, and RPA14. Phosphorylation of the N-terminal region of the RPA32 subunit plays an essential role in DNA metabolism in processes such as replication and damage response. Phosphorylated RPA32 (pRPA32) binds to RPA70 and possibly regulates the transient RPA70-Bloom syndrome helicase (BLM) interaction to inhibit DNA resection. Read More

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December 2020

LIM Protein Ajuba associates with the RPA complex through direct cell cycle-dependent interaction with the RPA70 subunit.

Sci Rep 2018 06 22;8(1):9536. Epub 2018 Jun 22.

Department of Biological Sciences, Hunter College, 695 Park Avenue, New York, NY, 10065, USA.

DNA damage response pathways are essential for genome stability and cell survival. Specifically, the ATR kinase is activated by DNA replication stress. An early event in this activation is the recruitment and phosphorylation of RPA, a single stranded DNA binding complex composed of three subunits, RPA70, RPA32 and RPA14. Read More

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RPA binds histone H3-H4 and functions in DNA replication-coupled nucleosome assembly.

Science 2017 01;355(6323):415-420

State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences and Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China.

DNA replication-coupled nucleosome assembly is essential to maintain genome integrity and retain epigenetic information. Multiple involved histone chaperones have been identified, but how nucleosome assembly is coupled to DNA replication remains elusive. Here we show that replication protein A (RPA), an essential replisome component that binds single-stranded DNA, has a role in replication-coupled nucleosome assembly. Read More

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January 2017

Metal binding mediated conformational change of XPA protein:a potential cytotoxic mechanism of nickel in the nucleotide excision repair.

J Mol Model 2016 07 16;22(7):156. Epub 2016 Jun 16.

Department of Pharmaceutical Sciences and Computational Chemical Genomics Screening Center, School of Pharmacy; NIH National Center of Excellence for Computational Drug Abuse Research; Drug Discovery Institute; Department of Computational Biology and Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15260, USA.

Nucleotide excision repair (NER) is a pivotal life process for repairing DNA nucleotide mismatch caused by chemicals, metal ions, radiation, and other factors. As the initiation step of NER, the xeroderma pigmentosum complementation group A protein (XPA) recognizes damaged DNA molecules, and recruits the replication protein A (RPA), another important player in the NER process. The stability of the Zn(2+)-chelated Zn-finger domain of XPA center core portion (i. Read More

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Interaction pattern for the complex of B-DNA Fullerene compounds with a set of known replication proteins using docking study.

Bioinformation 2015 31;11(3):122-6. Epub 2015 Mar 31.

Departments of Biosciences, Integral University, Lucknow, India.

Fullerenes have attracted considerable attention due to their unique chemical structure and potential applications which has opened wide venues for possible human exposure to various fullerene types. Therefore, in depth knowledge of how fullerene may interfere with various cellular processes becomes quite imperative. The present study was designed to investigate how the presence of fullerene affect the binding of DNA with different enzymes involved in replication process. Read More

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Blocking single-stranded transferred DNA conversion to double-stranded intermediates by overexpression of yeast DNA REPLICATION FACTOR A.

Plant Physiol 2015 Jan 25;167(1):153-63. Epub 2014 Nov 25.

Golan Research Institute, University of Haifa, Qatzrin 12900, Israel (M.D.-Y., R.D.);Noga AgroTech Desert Agriculture, Kmehin 85511, Israel (A.L.);Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109 (R.D., T.T.); andDepartment of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva 84105, Israel (T.T.).

Agrobacterium tumefaciens delivers its single-stranded transferred DNA (T-strand) into the host cell nucleus, where it can be converted into double-stranded molecules. Various studies have revealed that double-stranded transfer DNA (T-DNA) intermediates can serve as substrates by as yet uncharacterized integration machinery. Nevertheless, the possibility that T-strands are themselves substrates for integration cannot be ruled out. Read More

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January 2015

RPA antagonizes microhomology-mediated repair of DNA double-strand breaks.

Nat Struct Mol Biol 2014 Apr 9;21(4):405-12. Epub 2014 Mar 9.

Department of Microbiology and Immunology, Columbia University College of Physicians and Surgeons, New York, New York, USA.

Microhomology-mediated end joining (MMEJ) is a Ku- and ligase IV-independent mechanism for the repair of DNA double-strand breaks that contributes to chromosome rearrangements. Here we used a chromosomal end-joining assay to determine the genetic requirements for MMEJ in Saccharomyces cerevisiae. We found that end resection influences the ability to expose microhomologies; however, it is not rate limiting for MMEJ in wild-type cells. Read More

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Rpa43 and its partners in the yeast RNA polymerase I transcription complex.

FEBS Lett 2011 Nov 4;585(21):3355-9. Epub 2011 Oct 4.

CEA, iBiTec-S, Service de Biologie Intégrative & Génétique Moléculaire, F-91191 Gif-sur-Yvette, France.

An Rpa43/Rpa14 stalk protrudes from RNA polymerase I (RNAPI), with homology to Rpb7/Rpb4 (RNAPII), Rpc25/Rpc17 (RNAPIII) and RpoE/RpoF (archaea). In fungi and vertebrates, Rpa43 contains hydrophilic domains forming about half of its size, but these domains lack in Schizosaccharomyces pombe and most other eukaryote lineages. In Saccharomyces cerevisiae, they can be lost with little or no growth effect, as shown by deletion mapping and by domain swapping with fission yeast, but genetically interact with rpa12Δ, rpa34Δ or rpa49Δ, lacking non-essential subunits important for transcript elongation. Read More

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November 2011

Transcription by the multifunctional RNA polymerase I in Trypanosoma brucei functions independently of RPB7.

Mol Biochem Parasitol 2011 Nov 23;180(1):35-42. Epub 2011 Jul 23.

Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, CT 06030-6403, USA.

Trypanosoma brucei has a multifunctional RNA polymerase (pol) I that transcribes ribosomal gene units (RRNA) and units encoding its major cell surface proteins variant surface glycoprotein (VSG) and procyclin. Previous analysis of tandem affinity-purified, transcriptionally active RNA pol I identified ten subunits including an apparently trypanosomatid-specific protein termed RPA31. Another ortholog was identified in silico. Read More

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November 2011

Structural bases of dimerization of yeast telomere protein Cdc13 and its interaction with the catalytic subunit of DNA polymerase α.

Cell Res 2011 Feb 28;21(2):258-74. Epub 2010 Sep 28.

Howard Hughes Medical Institute, University of Michigan Medical School, 1150 W. Medical Center Drive, Ann Arbor, MI 48109, USA.

Budding yeast Cdc13-Stn1-Ten1 (CST) complex plays an essential role in telomere protection and maintenance, and has been proposed to be a telomere-specific replication protein A (RPA)-like complex. Previous genetic and structural studies revealed a close resemblance between Stn1-Ten1 and RPA32-RPA14. However, the relationship between Cdc13 and RPA70, the largest subunit of RPA, has remained unclear. Read More

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February 2011

Arabidopsis replication protein A 70a is required for DNA damage response and telomere length homeostasis.

Plant Cell Physiol 2009 Nov 6;50(11):1965-76. Epub 2009 Oct 6.

Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan.

Replication protein A1 (RPA1/RPA70) forms a heterotrimeric complex together with RPA2/RPA32 and RPA3/RPA14 subunits which plays essential roles in various aspects of DNA metabolism including replication, repair, recombination and telomere maintenance. Compared with RPA70 in yeast and mammals, limited information is available about the factor in plants. In this study, we analyzed the functions of AtRPA70a, which is most similar to human RPA70 among four paralogs in Arabidopsis thaliana. Read More

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November 2009

Structure of the full-length human RPA14/32 complex gives insights into the mechanism of DNA binding and complex formation.

J Mol Biol 2007 Dec 2;374(4):865-76. Epub 2007 Oct 2.

The Eppley Institute for Research in Cancer and Allied Diseases, 987696 Nebraska Medical Center, Omaha, NE 68198-7696, USA.

Replication protein A (RPA) is the ubiquitous, eukaryotic single-stranded DNA (ssDNA) binding protein and is essential for DNA replication, recombination, and repair. Here, crystal structures of the soluble RPA heterodimer, composed of the RPA14 and RPA32 subunits, have been determined for the full-length protein in multiple crystal forms. In all crystals, the electron density for the N-terminal (residues 1-42) and C-terminal (residues 175-270) regions of RPA32 is weak and of poor quality indicating that these regions are disordered and/or assume multiple positions in the crystals. Read More

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December 2007

Structural characterization of human RPA sequential binding to single-stranded DNA using ssDNA as a molecular ruler.

Biochemistry 2007 Jul 21;46(28):8226-33. Epub 2007 Jun 21.

Department of Biochemistry and Molecular Biology, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, Tennessee 37614, USA.

Human replication protein A (RPA), a heterotrimer composed of RPA70, RPA32, and RPA14 subunits, contains four single-stranded DNA (ssDNA) binding domains (DBD): DBD-A, DBD-B, and DBD-C in RPA70 and DBD-D in RPA32. Although crystallographic or NMR structures of these DBDs and a trimerization core have been determined, the structure of the full length of RPA or the RPA-ssDNA complex remains unknown. In this article, we have examined the structural features of RPA interaction with ssDNA by fluorescence spectroscopy. Read More

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Autoantibodies against the replication protein A complex in systemic lupus erythematosus and other autoimmune diseases.

Arthritis Res Ther 2006 ;8(4):R111

Division of Rheumatology and Clinical Immunology, Department of Medicine, University of Florida, PO Box 100221, Gainesville, Florida 32610, USA.

Replication protein A (RPA), a heterotrimer with subunits of molecular masses 70, 32, and 14 kDa, is a single-stranded-DNA-binding factor involved in DNA replication, repair, and recombination. There have been only three reported cases of anti-RPA in systemic lupus erythematosus (SLE) and Sjögren syndrome (SjS). This study sought to clarify the clinical significance of autoantibodies against RPA. Read More

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February 2007

A higher plant has three different types of RPA heterotrimeric complex.

J Biochem 2006 Jan;139(1):99-104

Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510.

Replication protein A (RPA) is a protein complex composed of three subunits known as RPA70, RPA32, and RPA14. Generally, only one version of each of the three RPA genes is present in animals and yeast (with the exception of the human RPA32 ortholog). In rice (Oryza sativa L. Read More

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January 2006

Phosphorylation of replication protein A by S-phase checkpoint kinases.

DNA Repair (Amst) 2006 Mar 18;5(3):369-80. Epub 2006 Jan 18.

Department of Microbiology & Immunology, and the Witebsky Center for Microbial Pathogenesis & Immunology, University at Buffalo, School of Medicine & Biomedical Sciences, Buffalo, NY 14214, USA.

The major eukaryotic single-stranded DNA (ssDNA) binding protein, replication protein A (RPA), is a heterotrimer with subunits of 70, 32 and 14 kDa (RPA70, RPA32 and RPA14). RPA-coated ssDNA has been implicated as one of the triggers for intra-S-phase checkpoint activation. Phosphorylation of RPA occurs in cells with damaged DNA or stalled replication forks. Read More

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Modulation of replication protein A function by its hyperphosphorylation-induced conformational change involving DNA binding domain B.

J Biol Chem 2005 Sep 9;280(38):32775-83. Epub 2005 Jul 9.

Department of Biochemistry and Molecular Biology, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, Tennessee 37614, USA.

Human replication protein A (RPA), composed of RPA70, RPA32, and RPA14 subunits, undergoes hyperphosphorylation in cells in response to DNA damage. Hyperphosphorylation that occurs predominately in the N-terminal region of RPA32 is believed to play a role in modulating the cellular activities of RPA essential for almost all DNA metabolic pathways. To understand how the hyperphosphorylation modulates the functions of RPA, we compared the structural characteristics of full-length native and hyperphosphorylated RPAs using mass spectrometric protein footprinting, fluorescence spectroscopy, and limited proteolysis. Read More

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September 2005

Interaction and colocalization of Rad9/Rad1/Hus1 checkpoint complex with replication protein A in human cells.

Oncogene 2005 Jul;24(29):4728-35

Department of Biochemistry and Molecular Biology, James H Quillen College of Medicine, East Tennessee State University, Johnson City, TN 37614, USA.

Replication protein A (RPA) is a eukaryotic single-stranded DNA-binding protein consisting of three subunits of 70-, 32-, and 14-kDa (RPA70, RPA32, RPA14, respectively). It is a protein essential for most cellular DNA metabolic pathways. Checkpoint proteins Rad9, Rad1, and Hus1 form a clamp-like complex which plays a central role in the DNA damage-induced checkpoint response. Read More

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Radiosensitivity is predicted by DNA end-binding complex density, but not by nuclear levels of band components.

Radiother Oncol 2004 Sep;72(3):325-32

Department of Experimental Radiation Oncology, University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 97, Houston, TX 77030, USA.

Background And Purpose: We previously determined that the density of a rapidly migrating DNA end-binding complex (termed 'band-A') predicts radiosensitivity of human normal and tumor cells. The goal of this study was first to identify the protein components of band-A and to determine if the protein levels of band-A components would correlate with band-A density and radiosensitivity.

Patients And Methods: DNA end-binding protein complex (DNA-EBC) protein components were identified by adding antibodies specific for a variety of DNA repair-associated proteins to the DNA-EBC reaction and then noting which antibodies super-shifted various DNA-EBC bands. Read More

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September 2004

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

J Biochem Mol Biol 2002 Mar;35(2):194-8

Department of Chemistry and Chemistry Institute for Functional Materials, Pusan National University, Pusan 609-735, Korea.

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). Read More

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Structure of the RPA trimerization core and its role in the multistep DNA-binding mechanism of RPA.

EMBO J 2002 Apr;21(7):1855-63

Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73190, USA.

The human single-stranded DNA-binding protein, replication protein A (RPA) binds DNA in at least two different modes: initial [8-10 nucleotides (nt)] and stable ( approximately 30 nt). Switching from 8 to 30 nt mode is associated with a large conformational change. Here we report the 2. Read More

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Rpa12p, a conserved RNA polymerase I subunit with two functional domains.

Mol Microbiol 2002 Mar;43(5):1105-13

Laboratoire de Génétique Moléculaire, URBM, Facultés Universitaires Notre-Dame de la Paix, 61 rue de Bruxelles, B-5000 Namur, Belgium.

Rpa12p is a subunit of RNA polymerase I formed of two zinc-binding domains. The N-terminal zinc region (positions 1-60) is poorly conserved from yeast to man. The C-terminal domain contains an invariant Q. Read More

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Replication protein A in Pyrococcus furiosus is involved in homologous DNA recombination.

Authors:
K Komori Y Ishino

J Biol Chem 2001 Jul 7;276(28):25654-60. Epub 2001 May 7.

Department of Molecular Biology, Biomolecular Engineering Research Institute, Suita, Osaka 565-0874, Japan.

Single-stranded DNA-binding protein in Bacteria and replication protein A (RPA) in Eukarya play crucial roles in DNA replication, repair, and recombination processes. We identified an RPA complex from the hyperthermophilic archaeon, Pyrococcus furiosus. Unlike the single-peptide RPAs from the methanogenic archaea, Methanococcus jannaschii and Methanothermobacter thermoautotrophicus, P. Read More

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Dynamic light-scattering analysis of full-length human RPA14/32 dimer: purification, crystallization and self-association.

Acta Crystallogr D Biol Crystallogr 2001 Feb;57(Pt 2):254-9

University of Toledo, Department of Chemistry, 2801 West Bancroft Street, Toledo, OH 43606, USA.

Replication protein A (RPA) is a single-stranded DNA-binding protein involved in all aspects of eukaryotic DNA metabolism. A soluble heterodimeric form of RPA is composed of 14 and 32 kDa subunits (RPA14/32). Dynamic light-scattering (DLS) analysis was used to improve the purification, stabilization and crystallization of RPA14/32. Read More

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February 2001

Structural basis for the recognition of DNA repair proteins UNG2, XPA, and RAD52 by replication factor RPA.

Cell 2000 Oct;103(3):449-56

Department of Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, USA.

Replication protein A (RPA), the nuclear ssDNA-binding protein in eukaryotes, is essential to DNA replication, recombination, and repair. We have shown that a globular domain at the C terminus of subunit RPA32 contains a specific surface that interacts in a similar manner with the DNA repair enzyme UNG2 and repair factors XPA and RAD52, each of which functions in a different repair pathway. NMR structures of the RPA32 domain, free and in complex with the minimal interaction domain of UNG2, were determined, defining a common structural basis for linking RPA to the nucleotide excision, base excision, and recombinational pathways of repairing damaged DNA. Read More

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October 2000

The role for zinc in replication protein A.

J Biol Chem 2000 Sep;275(35):27332-8

Department of Biochemistry and Molecular Biology, the University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73190, USA.

Heterotrimeric human single-stranded DNA (ssDNA)-binding protein, replication protein A (RPA), is a central player in DNA replication, recombination, and repair. The C terminus of the largest subunit, RPA70, contains a putative zinc-binding motif and is implicated in complex formation with two smaller subunits, RPA14 and RPA32. The C-terminal domain of RPA70 (RPA70-CTD) was characterized using proteolysis and x-ray fluorescence emission spectroscopy. Read More

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September 2000

Human replication protein A: global fold of the N-terminal RPA-70 domain reveals a basic cleft and flexible C-terminal linker.

J Biomol NMR 1999 Aug;14(4):321-31

Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA.

Human Replication Protein A (hsRPA) is required for multiple cellular processes in DNA metabolism including DNA repair, replication and recombination. It binds single-stranded DNA with high affinity and interacts specifically with multiple proteins. hsRPA forms a heterotrimeric complex composed of 70-, 32- and 14-kDa subunits (henceforth RPA70, RPA32, and RPA14). Read More

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The crystal structure of the complex of replication protein A subunits RPA32 and RPA14 reveals a mechanism for single-stranded DNA binding.

EMBO J 1999 Aug;18(16):4498-504

Department of Medical Genetics and Microbiology, University of Toronto, 1 King's College Circle, Toronto, Ontario M5S 1A8.

Replication protein A (RPA), the eukaryote single-stranded DNA-binding protein (SSB), is a heterotrimer. The largest subunit, RPA70, which harbours the major DNA-binding activity, has two DNA-binding domains that each adopt an OB-fold. The complex of the two smaller subunits, RPA32 and RPA14, has weak DNA-binding activity but the mechanism of DNA binding is unknown. Read More

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Phosphorylation of replication protein A middle subunit (RPA32) leads to a disassembly of the RPA heterotrimer.

J Biol Chem 1999 May;274(22):15556-61

Department of Biology, Universität Konstanz, D-78457 Konstanz, Germany.

Replication protein A (RPA), the major eukaryotic single-strand specific DNA binding protein, consists of three subunits, RPA70, RPA32, and RPA14. The middle subunit, RPA32, is phosphorylated in a cell cycle-dependent manner. RPA occurs in two nuclear compartments, bound to chromatin or free in the nucleosol. Read More

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