The splicing factor XAB2 interacts with ERCC1-XPF and XPG for R-loop processing

Evi Goulielmaki; Maria Tsekrekou; Nikos Batsiotos; Mariana Ascensão-Ferreira; Eleftheria Ledaki; Kalliopi Stratigi; Georgia Chatzinikolaou; Pantelis Topalis; Theodore Kosteas; Janine Altmüller; Jeroen A. Demmers; Nuno L. Barbosa-Morais; George A. Garinis

doi:10.1038/s41467-021-23505-1

The splicing factor XAB2 interacts with ERCC1-XPF and XPG for R-loop processing

Evi Goulielmaki
, Maria Tsekrekou
, Nikos Batsiotos
, Mariana Ascensão-Ferreira
, Eleftheria Ledaki
, Kalliopi Stratigi
, Georgia Chatzinikolaou
, Pantelis Topalis
, Theodore Kosteas
, Janine Altmüller
, Jeroen A. Demmers
, Nuno L. Barbosa-Morais
, George A. Garinis^*

^*Corresponding author for this work

Biochemistry

Foundation for Research and Technology-Hellas
University of Crete
Faculdade de Medicina da Universidade de Lisboa
University of Cologne

Research output: Contribution to journal › Article › Academic › peer-review

47 Citations (Scopus)

Abstract

RNA splicing, transcription and the DNA damage response are intriguingly linked in mammals but the underlying mechanisms remain poorly understood. Using an in vivo biotinylation tagging approach in mice, we show that the splicing factor XAB2 interacts with the core spliceosome and that it binds to spliceosomal U4 and U6 snRNAs and pre-mRNAs in developing livers. XAB2 depletion leads to aberrant intron retention, R-loop formation and DNA damage in cells. Studies in illudin S-treated cells and Csb^m/m developing livers reveal that transcription-blocking DNA lesions trigger the release of XAB2 from all RNA targets tested. Immunoprecipitation studies reveal that XAB2 interacts with ERCC1-XPF and XPG endonucleases outside nucleotide excision repair and that the trimeric protein complex binds RNA:DNA hybrids under conditions that favor the formation of R-loops. Thus, XAB2 functionally links the spliceosomal response to DNA damage with R-loop processing with important ramifications for transcription-coupled DNA repair disorders.

Original language	English
Article number	3153
Journal	Nature Communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-23505-1
Publication status	Published - 26 May 2021

Bibliographical note

Funding Information:
The Horizon 2020 ERC Consolidator grant “DeFiNER” (GA64663), the Horizon 2020 Marie Curie ITNs “Chromatin3D” (GA622934), “aDDRess” (GA812829) and “HealthAge” “(GA812830), the Hellenic Foundation for Research and Innovation (HFRI) and the General Secretariat for Research and Technology (GSRT) under grant agreement HFRI-1059 and HFRI-FM17-631 and the “Fondation Santé” grant supported this work. G.C. is supported by the IKY postdoctoral research fellowship program (MIS: 5001552), co-financed by the European Social Fund-ESF and the Greek government. N.L.B.-M. is supported by an EMBO Installation Grant (3057) and an Investigador FCT (Fundação para a Ciência e a Tecnologia) Starting Grant (IF/00595/2014); M.A.-F. is supported by an FCT PhD fellowship (PD/BD/128283/ 2017) and Fundação AstraZeneca.

Publisher Copyright:
© 2021, The Author(s).

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Goulielmaki, E., Tsekrekou, M., Batsiotos, N., Ascensão-Ferreira, M., Ledaki, E., Stratigi, K., Chatzinikolaou, G., Topalis, P., Kosteas, T., Altmüller, J., Demmers, J. A., Barbosa-Morais, N. L., & Garinis, G. A. (2021). The splicing factor XAB2 interacts with ERCC1-XPF and XPG for R-loop processing. Nature Communications, 12(1), Article 3153. https://doi.org/10.1038/s41467-021-23505-1

@article{208d504881304362b17a2382b21a4ca4,

title = "The splicing factor XAB2 interacts with ERCC1-XPF and XPG for R-loop processing",

abstract = "RNA splicing, transcription and the DNA damage response are intriguingly linked in mammals but the underlying mechanisms remain poorly understood. Using an in vivo biotinylation tagging approach in mice, we show that the splicing factor XAB2 interacts with the core spliceosome and that it binds to spliceosomal U4 and U6 snRNAs and pre-mRNAs in developing livers. XAB2 depletion leads to aberrant intron retention, R-loop formation and DNA damage in cells. Studies in illudin S-treated cells and Csbm/m developing livers reveal that transcription-blocking DNA lesions trigger the release of XAB2 from all RNA targets tested. Immunoprecipitation studies reveal that XAB2 interacts with ERCC1-XPF and XPG endonucleases outside nucleotide excision repair and that the trimeric protein complex binds RNA:DNA hybrids under conditions that favor the formation of R-loops. Thus, XAB2 functionally links the spliceosomal response to DNA damage with R-loop processing with important ramifications for transcription-coupled DNA repair disorders.",

author = "Evi Goulielmaki and Maria Tsekrekou and Nikos Batsiotos and Mariana Ascens{\~a}o-Ferreira and Eleftheria Ledaki and Kalliopi Stratigi and Georgia Chatzinikolaou and Pantelis Topalis and Theodore Kosteas and Janine Altm{\"u}ller and Demmers, \{Jeroen A.\} and Barbosa-Morais, \{Nuno L.\} and Garinis, \{George A.\}",

note = "Funding Information: The Horizon 2020 ERC Consolidator grant “DeFiNER” (GA64663), the Horizon 2020 Marie Curie ITNs “Chromatin3D” (GA622934), “aDDRess” (GA812829) and “HealthAge” “(GA812830), the Hellenic Foundation for Research and Innovation (HFRI) and the General Secretariat for Research and Technology (GSRT) under grant agreement HFRI-1059 and HFRI-FM17-631 and the “Fondation Sant{\'e}” grant supported this work. G.C. is supported by the IKY postdoctoral research fellowship program (MIS: 5001552), co-financed by the European Social Fund-ESF and the Greek government. N.L.B.-M. is supported by an EMBO Installation Grant (3057) and an Investigador FCT (Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia) Starting Grant (IF/00595/2014); M.A.-F. is supported by an FCT PhD fellowship (PD/BD/128283/ 2017) and Funda{\c c}{\~a}o AstraZeneca. Publisher Copyright: {\textcopyright} 2021, The Author(s).",

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month = may,

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doi = "10.1038/s41467-021-23505-1",

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Goulielmaki, E, Tsekrekou, M, Batsiotos, N, Ascensão-Ferreira, M, Ledaki, E, Stratigi, K, Chatzinikolaou, G, Topalis, P, Kosteas, T, Altmüller, J, Demmers, JA, Barbosa-Morais, NL & Garinis, GA 2021, 'The splicing factor XAB2 interacts with ERCC1-XPF and XPG for R-loop processing', Nature Communications, vol. 12, no. 1, 3153. https://doi.org/10.1038/s41467-021-23505-1

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T1 - The splicing factor XAB2 interacts with ERCC1-XPF and XPG for R-loop processing

AU - Goulielmaki, Evi

AU - Tsekrekou, Maria

AU - Batsiotos, Nikos

AU - Ascensão-Ferreira, Mariana

AU - Ledaki, Eleftheria

AU - Stratigi, Kalliopi

AU - Chatzinikolaou, Georgia

AU - Topalis, Pantelis

AU - Kosteas, Theodore

AU - Altmüller, Janine

AU - Demmers, Jeroen A.

AU - Barbosa-Morais, Nuno L.

AU - Garinis, George A.

N1 - Funding Information: The Horizon 2020 ERC Consolidator grant “DeFiNER” (GA64663), the Horizon 2020 Marie Curie ITNs “Chromatin3D” (GA622934), “aDDRess” (GA812829) and “HealthAge” “(GA812830), the Hellenic Foundation for Research and Innovation (HFRI) and the General Secretariat for Research and Technology (GSRT) under grant agreement HFRI-1059 and HFRI-FM17-631 and the “Fondation Santé” grant supported this work. G.C. is supported by the IKY postdoctoral research fellowship program (MIS: 5001552), co-financed by the European Social Fund-ESF and the Greek government. N.L.B.-M. is supported by an EMBO Installation Grant (3057) and an Investigador FCT (Fundação para a Ciência e a Tecnologia) Starting Grant (IF/00595/2014); M.A.-F. is supported by an FCT PhD fellowship (PD/BD/128283/ 2017) and Fundação AstraZeneca. Publisher Copyright: © 2021, The Author(s).

PY - 2021/5/26

Y1 - 2021/5/26

N2 - RNA splicing, transcription and the DNA damage response are intriguingly linked in mammals but the underlying mechanisms remain poorly understood. Using an in vivo biotinylation tagging approach in mice, we show that the splicing factor XAB2 interacts with the core spliceosome and that it binds to spliceosomal U4 and U6 snRNAs and pre-mRNAs in developing livers. XAB2 depletion leads to aberrant intron retention, R-loop formation and DNA damage in cells. Studies in illudin S-treated cells and Csbm/m developing livers reveal that transcription-blocking DNA lesions trigger the release of XAB2 from all RNA targets tested. Immunoprecipitation studies reveal that XAB2 interacts with ERCC1-XPF and XPG endonucleases outside nucleotide excision repair and that the trimeric protein complex binds RNA:DNA hybrids under conditions that favor the formation of R-loops. Thus, XAB2 functionally links the spliceosomal response to DNA damage with R-loop processing with important ramifications for transcription-coupled DNA repair disorders.

AB - RNA splicing, transcription and the DNA damage response are intriguingly linked in mammals but the underlying mechanisms remain poorly understood. Using an in vivo biotinylation tagging approach in mice, we show that the splicing factor XAB2 interacts with the core spliceosome and that it binds to spliceosomal U4 and U6 snRNAs and pre-mRNAs in developing livers. XAB2 depletion leads to aberrant intron retention, R-loop formation and DNA damage in cells. Studies in illudin S-treated cells and Csbm/m developing livers reveal that transcription-blocking DNA lesions trigger the release of XAB2 from all RNA targets tested. Immunoprecipitation studies reveal that XAB2 interacts with ERCC1-XPF and XPG endonucleases outside nucleotide excision repair and that the trimeric protein complex binds RNA:DNA hybrids under conditions that favor the formation of R-loops. Thus, XAB2 functionally links the spliceosomal response to DNA damage with R-loop processing with important ramifications for transcription-coupled DNA repair disorders.

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DO - 10.1038/s41467-021-23505-1

M3 - Article

C2 - 34039990

AN - SCOPUS:85106959643

SN - 2041-1723

VL - 12

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 3153

ER -

The splicing factor XAB2 interacts with ERCC1-XPF and XPG for R-loop processing

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