TY - JOUR

T1 - Replication protein A as a modulator of the poly(ADP-ribose)polymerase 1 activity

AU - Maltseva, Ekaterina A.

AU - Krasikova, Yulia S.

AU - Sukhanova, Maria V.

AU - Rechkunova, Nadejda I.

AU - Lavrik, Olga I.

N1 - Publisher Copyright: © 2018 Elsevier B.V.

PY - 2018/12/1

Y1 - 2018/12/1

N2 - Replication protein A contributes to all major pathways of DNA metabolism and is a target for post-translation modifications, including poly(ADP-ribosyl)ation catalyzed by PARP1. Here we demonstrate that the efficiency of RPA poly(ADP-ribosyl)ation strongly depends on the structure of DNA used for PARP1 activation and on the polarity of RPA binding. Moreover, RPA influences PARP1 activity, and this effect also depends on DNA structure: RPA inhibits PAR synthesis catalyzed by PARP1 in the presence of ssDNA and stimulates it in the presence of a DNA duplex, in particular that containing a nick or a gap. Using fluorescently labeled proteins, we showed their direct interaction and characterized it quantitatively. RPA can accelerate the replacement of poly(ADP-ribosyl)ated PARP1 molecules bound to DNA by the unmodified ones. Thus, our data allow us to suggest that the balance between the affinities of PARP1 and RPA for DNA and the interaction of these proteins with each other are the cornerstone of the modulating effect of RPA on PARP1 activity. This effect might contribute to the regulation of PARP1 activity in various DNA processing mechanisms including DNA replication and repair pathways, where both PARP1 and RPA participate.

AB - Replication protein A contributes to all major pathways of DNA metabolism and is a target for post-translation modifications, including poly(ADP-ribosyl)ation catalyzed by PARP1. Here we demonstrate that the efficiency of RPA poly(ADP-ribosyl)ation strongly depends on the structure of DNA used for PARP1 activation and on the polarity of RPA binding. Moreover, RPA influences PARP1 activity, and this effect also depends on DNA structure: RPA inhibits PAR synthesis catalyzed by PARP1 in the presence of ssDNA and stimulates it in the presence of a DNA duplex, in particular that containing a nick or a gap. Using fluorescently labeled proteins, we showed their direct interaction and characterized it quantitatively. RPA can accelerate the replacement of poly(ADP-ribosyl)ated PARP1 molecules bound to DNA by the unmodified ones. Thus, our data allow us to suggest that the balance between the affinities of PARP1 and RPA for DNA and the interaction of these proteins with each other are the cornerstone of the modulating effect of RPA on PARP1 activity. This effect might contribute to the regulation of PARP1 activity in various DNA processing mechanisms including DNA replication and repair pathways, where both PARP1 and RPA participate.

KW - PARP1

KW - Poly(ADP-ribosyl)ation

KW - Protein–protein interaction

KW - RPA

KW - POLARITY

KW - DNA

KW - POLYMERASE

KW - EXCISION

KW - STRESS

KW - BINDING

KW - STRAND

KW - Protein protein interaction

KW - DNA/metabolism

KW - Poly Adenosine Diphosphate Ribose/metabolism

KW - Biocatalysis

KW - Humans

KW - Replication Protein A/metabolism

KW - Poly (ADP-Ribose) Polymerase-1/metabolism

KW - Protein Processing, Post-Translational

UR - http://www.scopus.com/inward/record.url?scp=85054149315&partnerID=8YFLogxK

U2 - 10.1016/j.dnarep.2018.09.010

DO - 10.1016/j.dnarep.2018.09.010

M3 - Article

C2 - 30291044

AN - SCOPUS:85054149315

VL - 72

SP - 28

EP - 38

JO - DNA Repair

JF - DNA Repair

SN - 1568-7864

ER -