Mutational analysis of a function of xeroderma pigmentosum group A (XPA) protein in strand-specific DNA repair

Kobayashi, Tetsuo; Takeuchi, Seiji; Saijo, Masafumi; Nakatsu, Yoshimichi; Morioka, Hiroshi; Otsuka, Eizo; Wakasugi, Mitsuo; Nikaido, Osamu; Tanaka, Kiyoji

doi:10.1093/nar/26.20.4662

Cited by 44 publications

(27 citation statements)

References 45 publications

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“…The repair of CPD was examined in the 17.9-kb KpnI fragment within the dihydrofolate reductase (DHFR) gene in XP4PASV cells transfected with control or BRCA1 siRNA and irradiated with 8 J ⁄ m 2 using a method previously described. (30) Briefly, DNA was extracted, digested with KpnI and treated with T4 endonuclease V, which generates single-strand breaks at CPD sites. The samples were separated by electrophoresis in 0.65% alkaline agarose gels, transferred onto Hybond N + membranes (Amersham Biosciences, Little Chalfont, Bucks, UK), and hybridized with strand-specific digoxigenin (DIG)-labeled DNA probes.…”

Section: Methodsmentioning

confidence: 99%

BRCA1 contributes to transcription‐coupled repair of DNA damage through polyubiquitination and degradation of Cockayne syndrome B protein

Wei

Lan²,

Yasui

et al. 2011

Cancer Science

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BRCA1 is an important gene involved in susceptibility to breast and ovarian cancer and its product regulates the cellular response to DNA double-strand breaks. Here, we present evidence that BRCA1 also contributes to the transcription-coupled repair (TCR) of ultraviolet (UV) light-induced DNA damage. BRCA1 immediately accumulates at the sites of UV irradiation-mediated damage in cell nuclei in a manner that is fully dependent on both Cockayne syndrome B (CSB) protein and active transcription. Suppression of BRCA1 expression inhibits the TCR of UV lesions and increases the UV sensitivity of cells proficient in TCR. BRCA1 physically interacts with CSB protein. BRCA1 polyubiquitinates CSB and this polyubiquitination and subsequent degradation of CSB occur following UV irradiation, even in the absence of Cockayne syndrome A (CSA) protein. The depletion of BRCA1 expression increases the UV sensitivity of CSA-deficient cells. These results indicate that BRCA1 is involved in TCR and that a BRCA1-dependent polyubiquitination pathway for CSB exists alongside the CSA-dependent pathway to yield more efficient excision repair of lesions on the transcribed DNA strand. (Cancer Sci 2011; 102: 1840-1847 B RCA1 is an important breast and ovarian cancer susceptibility gene.(1,2) BRCA1 mutations are rare in sporadic breast and ovarian cancers (3,4) and its expression in these cancers is often reduced, (5) suggesting that BRCA1 plays a role in both hereditary and sporadic carcinogenesis. BRCA1 contains a RING domain at the amino (N)-terminus and two BRCA1 carboxy-terminal (BRCT) domains at the carboxy (C)-terminus. RING domain is an essential component of many ubiquitine E3 ligase. BRCA1 associates with BARD1, which also has a RING domain, (6) and the BRCA1 ⁄ BARD1 heterodimer has ubiquitin ligase activity. (7)(8)(9) BRCA1 has been implicated in a variety of biological processes, including DNA repair, transcription, chromatin remodeling and centrosome duplication.(10) BRCA1 localizes to nuclear foci during S-phase of the cell cycle.(11) Various mediators of DNA damage such as ultraviolet (UV) irradiation disperse the BRCA1 foci, followed by the reappearance of BRCA1 foci. (12) BRCA1 is phosphorylated in response to UV-induced damage.(13) BRCA1 associates with RNA polymerase II (RNA-PII) (14) and mediates the ubiquitination of RNAPII following UV irradiation. (15)(16)(17) The main type of DNA damage induced by UV irradiation is the formation of cyclobutane pyrimidine dimers (CPD) and (6-4) photoproduct adducts. These lesions are removed by nucleotide excision repair (NER). The NER operates via two pathways: transcription-coupled repair (TCR) and global genome repair (GGR). The TCR efficiently removes DNA lesions on the transcribed strands of transcriptionally active genes, whereas the GGR repairs DNA lesions throughout the genome. A stalled RNAPII is presumed to trigger the initiation of TCR in harmony with Cockayne syndrome (CS) proteins. Xeroderma pigmentosum (XP) and CS are rare genetic disorders. Xeroderma pigmentosum is charact...

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Section: Methodsmentioning

confidence: 99%

BRCA1 contributes to transcription‐coupled repair of DNA damage through polyubiquitination and degradation of Cockayne syndrome B protein

Wei

Lan²,

Yasui

et al. 2011

Cancer Science

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“…To investigate the role of the XPA interaction with RPA32 in NER, we prepared and examined the repair activities of mutant XPA proteins with N-terminal deletions. We have reported that the XP-A transfectant expressing a mutant XPA protein with a deletion of the N-terminal 36 amino acid residues (N⌬36) was more sensitive to UV irradiation than the transfectant expressing wild-type XPA and that the repair of the CPD is decreased in N⌬36-expressing cells (31).…”

Section: Analysis Of Mutant Xpa Proteins With N-terminal Deletions-mentioning

confidence: 99%

Nucleotide Excision Repair by Mutant Xeroderma Pigmentosum Group A (XPA) Proteins with Deficiency in Interaction with RPA

Saijo

Takedachi

Tanaka

2011

Journal of Biological Chemistry

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The xeroderma pigmentosum group A protein (XPA) is a core component of nucleotide excision repair (NER). To coordinate early stage NER, XPA interacts with various proteins, including replication protein A (RPA), ERCC1, DDB2, and TFIIH, in addition to UV-damaged or chemical carcinogendamaged DNA. In this study, we investigated the effects of mutations in the RPA binding regions of XPA on XPA function in NER. XPA binds through an N-terminal region to the middle subunit (RPA32) of the RPA heterotrimer and through a central region that overlaps with its damaged DNA binding region to the RPA70 subunit. In cell-free NER assays, an N-terminal deletion mutant of XPA showed loss of binding to RPA32 and reduced DNA repair activity, but it could still bind to UV-damaged DNA and RPA. In contrast, amino acid substitutions in the central region reduced incisions at the damaged site in the cell-free NER assay, and four of these mutants (K141A, T142A, K167A, and K179A) showed reduced binding to RPA70 but normal binding to damaged DNA. Furthermore, mutants that had one of the four aforementioned substitutions and an Nterminal deletion exhibited lower DNA incision activity and binding to RPA than XPA with only one of these substitutions or the deletion. Taken together, these results indicate that XPA interaction with both RPA32 and RPA70 is indispensable for NER reactions.

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“…Several lines of evidence lead to the currently accepted hypothesis that the lesions are initially recognized by XPC with the help of UV-DDB (UV DNA damage binding protein) for cyclobutane pyrimidine dimer (CPD) lesions (17)(18)(19)(20) followed by damage verification by XPD/TFIIH (21)(22)(23)(24), XPA, and XPG (3) to finally assemble to the preincision complex (3,(25)(26)(27). This step seems to involve recruitment of the XPA protein, which is one of the proteins essential for both GG-NER and TC-NER (28). As such, mutations of the XPA proteins provide one of the strongest NER phenotypes (29).…”

mentioning

confidence: 99%

“…Initial structural insights into the protein architecture were derived from an NMR structure of the DNA binding domain of XPA (39,40). However, the precise structural basis of the interactions of XPA with its DNA substrates is not known (3,28).…”

mentioning

confidence: 99%

Structural insights into the recognition of cisplatin and AAF-dG lesion by Rad14 (XPA)

Koch

Kuper

Gasteiger

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Nucleotide excision repair (NER) is responsible for the removal of a large variety of structurally diverse DNA lesions. Mutations of the involved proteins cause the xeroderma pigmentosum (XP) cancer predisposition syndrome. Although the general mechanism of the NER process is well studied, the function of the XPA protein, which is of central importance for successful NER, has remained enigmatic. It is known, that XPA binds kinked DNA structures and that it interacts also with DNA duplexes containing certain lesions, but the mechanism of interactions is unknown. Here we present two crystal structures of the DNA binding domain (DBD) of the yeast XPA homolog Rad14 bound to DNA with either a cisplatin lesion (1,2-GG) or an acetylaminofluorene adduct (AAF-dG). In the structures, we see that two Rad14 molecules bind to the duplex, which induces DNA melting of the duplex remote from the lesion. Each monomer interrogates the duplex with a β-hairpin, which creates a 13mer duplex recognition motif additionally characterized by a sharp 70°DNA kink at the position of the lesion. Although the 1,2-GG lesion stabilizes the kink due to the covalent fixation of the crosslinked dG bases at a 90°angle, the AAF-dG fully intercalates into the duplex to stabilize the kinked structure.

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Mutational analysis of a function of xeroderma pigmentosum group A (XPA) protein in strand-specific DNA repair

Cited by 44 publications

References 45 publications

BRCA1 contributes to transcription‐coupled repair of DNA damage through polyubiquitination and degradation of Cockayne syndrome B protein

BRCA1 contributes to transcription‐coupled repair of DNA damage through polyubiquitination and degradation of Cockayne syndrome B protein

Nucleotide Excision Repair by Mutant Xeroderma Pigmentosum Group A (XPA) Proteins with Deficiency in Interaction with RPA

Structural insights into the recognition of cisplatin and AAF-dG lesion by Rad14 (XPA)

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