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Functional Roles of PARP2 in Assembling Protein–Protein Complexes Involved in Base Excision DNA Repair

Poly(ADP-ribose) polymerase 2 (PARP2) participates in base excision repair (BER) alongside PARP1, but its functions are still under study. Here, we characterize binding affinities of PARP2 for other BER proteins (PARP1, APE1, Polβ, and XRCC1) and oligomerization states of the homo- and hetero-associ...

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Autores principales: Vasil’eva, Inna, Moor, Nina, Anarbaev, Rashid, Kutuzov, Mikhail, Lavrik, Olga
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8124814/
https://www.ncbi.nlm.nih.gov/pubmed/33925170
http://dx.doi.org/10.3390/ijms22094679
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author Vasil’eva, Inna
Moor, Nina
Anarbaev, Rashid
Kutuzov, Mikhail
Lavrik, Olga
author_facet Vasil’eva, Inna
Moor, Nina
Anarbaev, Rashid
Kutuzov, Mikhail
Lavrik, Olga
author_sort Vasil’eva, Inna
collection PubMed
description Poly(ADP-ribose) polymerase 2 (PARP2) participates in base excision repair (BER) alongside PARP1, but its functions are still under study. Here, we characterize binding affinities of PARP2 for other BER proteins (PARP1, APE1, Polβ, and XRCC1) and oligomerization states of the homo- and hetero-associated complexes using fluorescence-based and light scattering techniques. To compare PARP2 and PARP1 in the efficiency of PAR synthesis, in the absence and presence of protein partners, the size of PARP2 PARylated in various reaction conditions was measured. Unlike PARP1, PARP2 forms more dynamic complexes with common protein partners, and their stability is effectively modulated by DNA intermediates. Apparent binding affinity constants determined for homo- and hetero-oligomerized PARP1 and PARP2 provide evidence that the major form of PARP2 at excessive PARP1 level is their heterocomplex. Autoregulation of PAR elongation at high PARP and NAD(+) concentrations is stronger for PARP2 than for PARP1, and the activity of PARP2 is more effectively inhibited by XRCC1. Moreover, the activity of both PARP1 and PARP2 is suppressed upon their heteroPARylation. Taken together, our findings suggest that PARP2 can function differently in BER, promoting XRCC1-dependent repair (similarly to PARP1) or an alternative XRCC1-independent mechanism via hetero-oligomerization with PARP1.
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spelling pubmed-81248142021-05-17 Functional Roles of PARP2 in Assembling Protein–Protein Complexes Involved in Base Excision DNA Repair Vasil’eva, Inna Moor, Nina Anarbaev, Rashid Kutuzov, Mikhail Lavrik, Olga Int J Mol Sci Article Poly(ADP-ribose) polymerase 2 (PARP2) participates in base excision repair (BER) alongside PARP1, but its functions are still under study. Here, we characterize binding affinities of PARP2 for other BER proteins (PARP1, APE1, Polβ, and XRCC1) and oligomerization states of the homo- and hetero-associated complexes using fluorescence-based and light scattering techniques. To compare PARP2 and PARP1 in the efficiency of PAR synthesis, in the absence and presence of protein partners, the size of PARP2 PARylated in various reaction conditions was measured. Unlike PARP1, PARP2 forms more dynamic complexes with common protein partners, and their stability is effectively modulated by DNA intermediates. Apparent binding affinity constants determined for homo- and hetero-oligomerized PARP1 and PARP2 provide evidence that the major form of PARP2 at excessive PARP1 level is their heterocomplex. Autoregulation of PAR elongation at high PARP and NAD(+) concentrations is stronger for PARP2 than for PARP1, and the activity of PARP2 is more effectively inhibited by XRCC1. Moreover, the activity of both PARP1 and PARP2 is suppressed upon their heteroPARylation. Taken together, our findings suggest that PARP2 can function differently in BER, promoting XRCC1-dependent repair (similarly to PARP1) or an alternative XRCC1-independent mechanism via hetero-oligomerization with PARP1. MDPI 2021-04-28 /pmc/articles/PMC8124814/ /pubmed/33925170 http://dx.doi.org/10.3390/ijms22094679 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Vasil’eva, Inna
Moor, Nina
Anarbaev, Rashid
Kutuzov, Mikhail
Lavrik, Olga
Functional Roles of PARP2 in Assembling Protein–Protein Complexes Involved in Base Excision DNA Repair
title Functional Roles of PARP2 in Assembling Protein–Protein Complexes Involved in Base Excision DNA Repair
title_full Functional Roles of PARP2 in Assembling Protein–Protein Complexes Involved in Base Excision DNA Repair
title_fullStr Functional Roles of PARP2 in Assembling Protein–Protein Complexes Involved in Base Excision DNA Repair
title_full_unstemmed Functional Roles of PARP2 in Assembling Protein–Protein Complexes Involved in Base Excision DNA Repair
title_short Functional Roles of PARP2 in Assembling Protein–Protein Complexes Involved in Base Excision DNA Repair
title_sort functional roles of parp2 in assembling protein–protein complexes involved in base excision dna repair
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8124814/
https://www.ncbi.nlm.nih.gov/pubmed/33925170
http://dx.doi.org/10.3390/ijms22094679
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