title: Human replication protein A. The C-terminal RPA70 and the central RPA32 domains are involved in the interactions with the 3'-end of a primer-template DNA.
abstract: Although the mechanical aspects of the single-stranded DNA (ssDNA) binding activity of human replication protein A (RPA) have been extensively studied, only limited information is available about its interaction with other physiologically relevant DNA structures. RPA interacts with partial DNA duplexes that resemble DNA intermediates found in the processes of DNA replication and DNA repair. Limited proteolysis of RPA showed that RPA associated with ssDNA is less protected against proteases than RPA bound to a partial duplex DNA containing a 5'-protruding tail that had the same length as the ssDNA. Modification of both the 70- and 32-kDa subunits, RPA70 and RPA32, respectively, by photoaffinity labeling indicates that RPA can bind the primer-template junction of partial duplex DNAs by interacting with the 3'-end of the primer. The identification of the protein domains modified by the photoreactive 3'-end of the primer showed that domains located in the central part of the RPA32 subunit (amino acids 39-180) and the C-terminal part of the RPA70 subunit (amino acids 432-616) are involved in these interactions.
authors.affiliation: Abteilung Biochemie, Institut für Molekulare Biotechnologie, D-07745 Jena, Germany.
authors.forename: Pavel E
authors.initials: PE
authors.lastname: Pestryakov
authors.forename: Klaus
authors.initials: K
authors.lastname: Weisshart
authors.forename: Bernhard
authors.initials: B
authors.lastname: Schlott
authors.forename: Svetlana N
authors.initials: SN
authors.lastname: Khodyreva
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kremmer
authors.forename: Frank
authors.initials: F
authors.lastname: Grosse
authors.forename: Olga I
authors.initials: OI
authors.lastname: Lavrik
authors.forename: Heinz-Peter
authors.initials: HP
authors.lastname: Nasheuer
mesh_terms: D024509:3' Flanking Region; D001665:Binding Sites; D017931:DNA Primers / Q000235:genetics / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051716:Replication Protein A
chemical_list: D017931:DNA Primers; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; C497167:RPA1 protein, human; D051716:Replication Protein A