Bernadó, Pau

Blanco, Francisco J.

Castillo, Francisco

Cordeiro, Tiago N.

De Biasio, Alfredo

de Opakua, Alain Ibáñez

Delgado, Sandra

Diercks, Tammo

Gil-Cartón, David

Luque, Irene

Merino, Nekane

Molina, Rafael

Montoya, Guillermo

Mortuza, Gulnahar B.

Villate, Maider

Nature Communications

English

The intrinsically disordered protein p15(PAF) regulates DNA replication and repair by binding to the proliferating cell nuclear antigen (PCNA) sliding clamp. We present the structure of the human p15(PAF)-PCNA complex. Crystallography and NMR show the central PCNA-interacting protein motif (PIP-box) of p15(PAF) tightly bound to the front-face of PCNA. In contrast to other PCNA-interacting proteins, p15(PAF) also contacts the inside of, and passes through, the PCNA ring. The disordered p15(PAF) termini emerge at opposite faces of the ring, but remain protected from 20S proteasomal degradation. Both free and PCNA-bound p15(PAF) binds DNA mainly through its histone-like N-terminal tail, while PCNA does not, and a model of the ternary complex with DNA inside the PCNA ring is consistent with electron micrographs. We propose that p15(PAF) acts as a flexible drag that regulates PCNA sliding along the DNA and facilitates the switch from replicative to translesion synthesis polymerase binding.</p

Mortuza, Gulnahar B

Cordeiro, Tiago N

Blanco, Francisco J

Copenhagen University Research Information System

Structure of p15<sup>PAF</sup>-PCNA complex and implications for clamp sliding during DNA replication and repair

The intrinsically disordered protein p15(PAF) regulates DNA replication and repair by binding to the proliferating cell nuclear antigen (PCNA) sliding clamp. We present the structure of the human p15(PAF)-PCNA complex. Crystallography and NMR show the central PCNA-interacting protein motif (PIP-box) of p15(PAF) tightly bound to the front-face of PCNA. In contrast to other PCNA-interacting proteins, p15(PAF) also contacts the inside of, and passes through, the PCNA ring. The disordered p15(PAF) termini emerge at opposite faces of the ring, but remain protected from 20S proteasomal degradation. Both free and PCNA-bound p15(PAF) binds DNA mainly through its histone-like N-terminal tail, while PCNA does not, and a model of the ternary complex with DNA inside the PCNA ring is consistent with electron micrographs. We propose that p15(PAF) acts as a flexible drag that regulates PCNA sliding along the DNA and facilitates the switch from replicative to translesion synthesis polymerase binding.SíWe thank P. Redondo for help with the DNA duplex formation, T. Kaminishi for help with molecular modelling, M. Lelli for help with NMR relaxation measurements, S. Pongor for help with fluorescence polarization, N. Mailand for pointing out the p15 sequence similarity with histone H3 and S. Onesti for helpful discussions. This work was supported by the Spanish MINECO grants CTQ2011-28680 to F.J.B., BFU2011-23815 to G.M., and Juan de la Cierva to A. De. B., grant NNF14CC0001 to G.M., and SPIN-HD-ANR Chaires d'Excellence and the ATIP-Avenir to P. B. This research received funding from the European Community's FP7 grant agreement number 283570 (BioStruct-X, providing acces to SLS and ALBA syncrotrons) and 261863 (Bio-NMR, providing access to the 1 GHz NMR spectrometer at the Rhone-Alpes European Large Scale Facility for NMR)

REPISALUD (Instituto de Salud Carlos III)

Structure of p15(PAF)-PCNA complex and implications for clamp sliding during DNA replication and repair

12 pages, 6 figures, 1 tableThe intrinsically disordered protein p15(PAF) regulates DNA replication and repair by binding to the proliferating cell nuclear antigen (PCNA) sliding clamp. We present the structure of the human p15(PAF)-PCNA complex. Crystallography and NMR show the central PCNA-interacting protein motif (PIP-box) of p15(PAF) tightly bound to the front-face of PCNA. In contrast to other PCNA-interacting proteins, p15(PAF) also contacts the inside of, and passes through, the PCNA ring. The disordered p15(PAF) termini emerge at opposite faces of the ring, but remain protected from 20S proteasomal degradation. Both free and PCNA-bound p15(PAF) binds DNA mainly through its histone-like N-terminal tail, while PCNA does not, and a model of the ternary complex with DNA inside the PCNA ring is consistent with electron micrographs. We propose that p15(PAF) acts as a flexible drag that regulates PCNA sliding along the DNA and facilitates the switch from replicative to translesion synthesis polymerase binding.This work was supported by the Spanish MINECO grants CTQ2011-28680 to F.J.B., BFU2011-23815 to G.M., and Juan de la Cierva to A.De.B., grant NNF14CC0001 to G.M., and SPIN-HD–ANR Chaires d’Excellence and the ATIP-Avenir to P.B. This research received funding from the European Community’s FP7 grant agreement number 283570 (BioStruct-X, providing acces to SLS and ALBA syncrotrons) and 261863 (Bio-NMR, providing access to the 1 GHz NMR spectrometer at the Rhône-Alpes European Large Scale Facility for NMR).Peer reviewe

Biasio, Alfredo de

Ibáñez de Opakua, Alain

Digital.CSIC

Alfredo De Biasio

Alain Ibáñez de Opakua

Gulnahar B. Mortuza

Rafael Molina

Tiago N. Cordeiro

Francisco Castillo

Maider Villate

Nekane Merino

Sandra Delgado

David Gil-Cartón

Irene Luque

Tammo Diercks

Pau Bernadó

Guillermo Montoya

Francisco J. Blanco

Crossref

Structure of p15PAF–PCNA complex and implications for clamp sliding during DNA replication and repair

https://www.openaccessrepository.it/record/26509/files/fulltext.pdf

Structure of p15PAF–PCNA complex and implications for clamp sliding during DNA replication and repair

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Copenhagen University Research Information System

REPISALUD (Instituto de Salud Carlos III)

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