Phosphorylation of replication protein A (RPA) by Cdk2 and the checkpoint kinase ATR (ATM and Rad3 related) during replication fork stalling stabilizes the replisome, but how these modifications safeguard the fork is not understood. To address this question, we used single-molecule fiber analysis in cells expressing a phosphorylation-defective RPA2 subunit or lacking phosphatase activity toward...

Replication protein A (RPA) is a heterotrimeric protein complex required for a large number of DNA metabolic processes, including DNA replication and repair. An alternative form of RPA (aRPA) has been described in which the RPA2 subunit (the 32-kDa subunit of RPA and product of the RPA2 gene) of canonical RPA is replaced by a homologous subunit, RPA4. The normal function of aRPA is not known; h...

Replication protein A (RPA) is a heterotrimeric protein required for many DNA metabolic functions, including replication, recombination, and nucleotide excision repair (NER). We report the pre-steady-state kinetic analysis of RPA-binding DNA substrates using a stopped-flow assay to elucidate the kinetics of DNA damage recognition. The bimolecular association rate, k(on), for RPA binding to dupl...

Replication protein A (RPA) is a trimeric single-stranded DNA-binding protein complex involved in DNA replication, repair, and recombination. DNA damage induces phosphorylation of the RPA p34 subunit, and it has been speculated that this phosphorylation could contribute to the regulation of the DNA damage-induced S-phase checkpoint. To further examine this potential relationship, human cell lin...

Saccharomyces cerevisiae RAD51, RAD55, and RAD57 genes, required for genetic recombination and DNA double-strand-break repair, encode proteins homologous to one another and to the Escherichia coli RecA protein. Rad51 protein catalyzes the DNA strand-exchange reaction with a dependence on ATP and on the heterotrimeric single-strand DNA (ssDNA) binding factor replication protein A (RPA). By sever...

The risk perception attitude (RPA) framework was tested as a message tailoring strategy to encourage diabetes screening. Participants (N = 602) were first categorized into one of four RPA groups based on their diabetes risk and efficacy perceptions and then randomly assigned to receive a message that matched their RPA, mismatched their RPA, or a control message. Participants receiving a matched...

Low molecular size additives such as L-arginine and the redox compounds have been used both in the culturemedium and in vitro refolding to increase recombinant proteins production. Additives increase proteinrefolding and yield of active proteins by suppressing aggregate formation or enhancing refolding process.In this work, a comparative study was performed on refolding of rec...

The serological response and efficacy of Bacillus anthracis recombinant protective antigen (rPA) vaccines formulated with aluminum hydroxide adjuvant, either with or without formaldehyde, were evaluated in rabbits. Rabbits that had been injected with a single dose of 25 microg of rPA adsorbed to 500 microg of aluminum in aluminum hydroxide gel (Alhydrogel) had a significantly higher quantitativ...

Processing of G4 DNA by Dna2 Helicase/Nuclease and RPA Provides Insights into the Mechanism of Dna2/RPA Substrate Recognition Taro Masuda-Sasa, Piotr Polaczek, Xiao P. Peng, Lu Chen and Judith L. Campbell Braun Laboratories, 147-75, California Institute of Technology, Pasadena, CA 91125 Running title: Dna2/RPA/G4 interactions

Three immunizations of mice with recombinant protective antigen (rPA) by transcutaneous immunization (TCI) induced long-term neutralizing antibody titers that were superior to those obtained with aluminum-adsorbed rPA. In addition, rPA alone exhibited adjuvant activity for TCI. Forty-six weeks after completion of TCI, 100% protection was observed against lethal anthrax challenge.