hSSB2 (NABP1) is required for the recruitment of RPA during the cellular response to DNA UV damage

Didier Boucher; Ruvini Kariawasam; Joshua Burgess; Adrian Gimenez; Tristan E. Ocampo; Blake Ferguson; Ali Naqi; Graeme J. Walker; Emma Bolderson; Roland Gamsjaeger; Kenneth J. O’Byrne; Liza Cubeddu; Kum Kum Khanna; Derek J. Richard

doi:10.1038/s41598-021-99355-0

hSSB2 (NABP1) is required for the recruitment of RPA during the cellular response to DNA UV damage

Didier Boucher, Ruvini Kariawasam, Joshua Burgess, Adrian Gimenez, Tristan E. Ocampo, Blake Ferguson, Ali Naqi, Graeme J. Walker, Emma Bolderson, Roland Gamsjaeger, Kenneth J. O’Byrne, Liza Cubeddu, Kum Kum Khanna, Derek J. Richard

Western Sydney University

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

Abstract

Maintenance of genomic stability is critical to prevent diseases such as cancer. As such, eukaryotic cells have multiple pathways to efficiently detect, signal and repair DNA damage. One common form of exogenous DNA damage comes from ultraviolet B (UVB) radiation. UVB generates cyclobutane pyrimidine dimers (CPD) that must be rapidly detected and repaired to maintain the genetic code. The nucleotide excision repair (NER) pathway is the main repair system for this type of DNA damage. Here, we determined the role of the human Single-Stranded DNA Binding protein 2, hSSB2, in the response to UVB exposure. We demonstrate that hSSB2 levels increase in vitro and in vivo after UVB irradiation and that hSSB2 rapidly binds to chromatin. Depletion of hSSB2 results in significantly decreased Replication Protein A (RPA32) phosphorylation and impaired RPA32 localisation to the site of UV-induced DNA damage. Delayed recruitment of NER protein Xeroderma Pigmentosum group C (XPC) was also observed, leading to increased cellular sensitivity to UVB. Finally, hSSB2 was shown to have affinity for single-strand DNA containing a single CPD and for duplex DNA with a two-base mismatch mimicking a CPD moiety. Altogether our data demonstrate that hSSB2 is involved in the cellular response to UV exposure.

Original language	English
Article number	20256
Number of pages	12
Journal	Scientific Reports
Volume	11
Issue number	1
DOIs	https://doi.org/10.1038/s41598-021-99355-0
Publication status	Published - 2021

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Boucher, D., Kariawasam, R., Burgess, J., Gimenez, A., Ocampo, T. E., Ferguson, B., Naqi, A., Walker, G. J., Bolderson, E., Gamsjaeger, R., O’Byrne, K. J., Cubeddu, L., Khanna, K. K., & Richard, D. J. (2021). hSSB2 (NABP1) is required for the recruitment of RPA during the cellular response to DNA UV damage. Scientific Reports, 11(1), Article 20256. https://doi.org/10.1038/s41598-021-99355-0

@article{255a5e1a95ea40d5adc44773fabf194f,

title = "hSSB2 (NABP1) is required for the recruitment of RPA during the cellular response to DNA UV damage",

abstract = "Maintenance of genomic stability is critical to prevent diseases such as cancer. As such, eukaryotic cells have multiple pathways to efficiently detect, signal and repair DNA damage. One common form of exogenous DNA damage comes from ultraviolet B (UVB) radiation. UVB generates cyclobutane pyrimidine dimers (CPD) that must be rapidly detected and repaired to maintain the genetic code. The nucleotide excision repair (NER) pathway is the main repair system for this type of DNA damage. Here, we determined the role of the human Single-Stranded DNA Binding protein 2, hSSB2, in the response to UVB exposure. We demonstrate that hSSB2 levels increase in vitro and in vivo after UVB irradiation and that hSSB2 rapidly binds to chromatin. Depletion of hSSB2 results in significantly decreased Replication Protein A (RPA32) phosphorylation and impaired RPA32 localisation to the site of UV-induced DNA damage. Delayed recruitment of NER protein Xeroderma Pigmentosum group C (XPC) was also observed, leading to increased cellular sensitivity to UVB. Finally, hSSB2 was shown to have affinity for single-strand DNA containing a single CPD and for duplex DNA with a two-base mismatch mimicking a CPD moiety. Altogether our data demonstrate that hSSB2 is involved in the cellular response to UV exposure.",

author = "Didier Boucher and Ruvini Kariawasam and Joshua Burgess and Adrian Gimenez and Ocampo, {Tristan E.} and Blake Ferguson and Ali Naqi and Walker, {Graeme J.} and Emma Bolderson and Roland Gamsjaeger and O{\textquoteright}Byrne, {Kenneth J.} and Liza Cubeddu and Khanna, {Kum Kum} and Richard, {Derek J.}",

year = "2021",

doi = "10.1038/s41598-021-99355-0",

language = "English",

volume = "11",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

number = "1",

}

Boucher, D, Kariawasam, R, Burgess, J, Gimenez, A, Ocampo, TE, Ferguson, B, Naqi, A, Walker, GJ, Bolderson, E, Gamsjaeger, R, O’Byrne, KJ, Cubeddu, L, Khanna, KK & Richard, DJ 2021, 'hSSB2 (NABP1) is required for the recruitment of RPA during the cellular response to DNA UV damage', Scientific Reports, vol. 11, no. 1, 20256. https://doi.org/10.1038/s41598-021-99355-0

TY - JOUR

T1 - hSSB2 (NABP1) is required for the recruitment of RPA during the cellular response to DNA UV damage

AU - Boucher, Didier

AU - Kariawasam, Ruvini

AU - Burgess, Joshua

AU - Gimenez, Adrian

AU - Ocampo, Tristan E.

AU - Ferguson, Blake

AU - Naqi, Ali

AU - Walker, Graeme J.

AU - Bolderson, Emma

AU - Gamsjaeger, Roland

AU - O’Byrne, Kenneth J.

AU - Cubeddu, Liza

AU - Khanna, Kum Kum

AU - Richard, Derek J.

PY - 2021

Y1 - 2021

N2 - Maintenance of genomic stability is critical to prevent diseases such as cancer. As such, eukaryotic cells have multiple pathways to efficiently detect, signal and repair DNA damage. One common form of exogenous DNA damage comes from ultraviolet B (UVB) radiation. UVB generates cyclobutane pyrimidine dimers (CPD) that must be rapidly detected and repaired to maintain the genetic code. The nucleotide excision repair (NER) pathway is the main repair system for this type of DNA damage. Here, we determined the role of the human Single-Stranded DNA Binding protein 2, hSSB2, in the response to UVB exposure. We demonstrate that hSSB2 levels increase in vitro and in vivo after UVB irradiation and that hSSB2 rapidly binds to chromatin. Depletion of hSSB2 results in significantly decreased Replication Protein A (RPA32) phosphorylation and impaired RPA32 localisation to the site of UV-induced DNA damage. Delayed recruitment of NER protein Xeroderma Pigmentosum group C (XPC) was also observed, leading to increased cellular sensitivity to UVB. Finally, hSSB2 was shown to have affinity for single-strand DNA containing a single CPD and for duplex DNA with a two-base mismatch mimicking a CPD moiety. Altogether our data demonstrate that hSSB2 is involved in the cellular response to UV exposure.

AB - Maintenance of genomic stability is critical to prevent diseases such as cancer. As such, eukaryotic cells have multiple pathways to efficiently detect, signal and repair DNA damage. One common form of exogenous DNA damage comes from ultraviolet B (UVB) radiation. UVB generates cyclobutane pyrimidine dimers (CPD) that must be rapidly detected and repaired to maintain the genetic code. The nucleotide excision repair (NER) pathway is the main repair system for this type of DNA damage. Here, we determined the role of the human Single-Stranded DNA Binding protein 2, hSSB2, in the response to UVB exposure. We demonstrate that hSSB2 levels increase in vitro and in vivo after UVB irradiation and that hSSB2 rapidly binds to chromatin. Depletion of hSSB2 results in significantly decreased Replication Protein A (RPA32) phosphorylation and impaired RPA32 localisation to the site of UV-induced DNA damage. Delayed recruitment of NER protein Xeroderma Pigmentosum group C (XPC) was also observed, leading to increased cellular sensitivity to UVB. Finally, hSSB2 was shown to have affinity for single-strand DNA containing a single CPD and for duplex DNA with a two-base mismatch mimicking a CPD moiety. Altogether our data demonstrate that hSSB2 is involved in the cellular response to UV exposure.

UR - https://hdl.handle.net/1959.7/uws:61724

U2 - 10.1038/s41598-021-99355-0

DO - 10.1038/s41598-021-99355-0

M3 - Article

SN - 2045-2322

VL - 11

JO - Scientific Reports

JF - Scientific Reports

IS - 1

M1 - 20256

ER -

hSSB2 (NABP1) is required for the recruitment of RPA during the cellular response to DNA UV damage

Abstract

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