Fate of RNA Polymerase II Stalled at a Cisplatin Lesion

Tremeau‐Bravard, Alexandre; Riedl, Thilo; Egly, Jean‐Marc; Dahmus, Michael E.

doi:10.1074/jbc.m309853200

Cited by 63 publications

(77 citation statements)

References 40 publications

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“…As a consequence, it is impossible to predict the mechanisms of transcriptional stalling or mutagenesis at other types of lesions. Instead, the detailed stalling mechanisms remain to be investigated for other lesions, including the less frequent 1,3-d(GpNpG) intrastrand platinum cross-link 6,7,18,19 . Nevertheless, this study and our previous study of CPD-induced transcriptional stalling 8 suggest general aspects of the recognition of bulky dinucleotide lesions by Pol II, including a translocation barrier between positions +2/+3 and +1/+2, an A-rule for nontemplated nucleotide incorporation, and the possibility of lesion bypass under conditions that circumvent the natural stalling mechanism.…”

Section: Discussionmentioning

confidence: 99%

Mechanism of transcriptional stalling at cisplatin-damaged DNA

Damsma

Alt

Brueckner

et al. 2007

Nat Struct Mol Biol

160

133

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The anticancer drug cisplatin forms 1,2-d(GpG) DNA intrastrand cross-links (cisplatin lesions) that stall RNA polymerase II (Pol II) and trigger transcription-coupled DNA repair. Here we present a structure-function analysis of Pol II stalling at a cisplatin lesion in the DNA template. Pol II stalling results from a translocation barrier that prevents delivery of the lesion to the active site. AMP misincorporation occurs at the barrier and also at an abasic site, suggesting that it arises from nontemplated synthesis according to an 'A-rule' known for DNA polymerases. Pol II can bypass a cisplatin lesion that is artificially placed beyond the translocation barrier, even in the presence of a G . A mismatch. Thus, the barrier prevents transcriptional mutagenesis. The stalling mechanism differs from that of Pol II stalling at a photolesion, which involves delivery of the lesion to the active site and lesion-templated misincorporation that blocks transcription.Cisplatin (cis-diamminedichloroplatinum(II)) is a widely used anticancer drug that forms DNA adducts that interfere with replication and transcription 1 . The most frequent cisplatin DNA adducts are 1,2-d(GpG) intrastrand cross-links, or cisplatin lesions, in which platinum coordinates the N7 atoms of adjacent guanosines in a DNA strand 2 (Fig. 1a). A cisplatin lesion in the DNA template strand blocks transcription elongation by the single-subunit RNA polymerase from phage T7 (ref.3) and by Pol II 4-6 , and leads to stable polymerase stalling 7 . The stalled Pol II elongation complex can be bound by the elongation factor TFIIS, which stimulates polymerase back-tracking and 3¢ RNA cleavage 6 . A small fraction of polymerases can apparently read through a cisplatin lesion 6 .The detailed molecular mechanisms of cisplatin DNA adduct processing by nucleic acid polymerases are not understood. Here we have used a combination of X-ray crystallography and RNA-extension assays to derive the molecular mechanism of Saccharomyces cerevisiae Pol II stalling at a cisplatin lesion. Comparison of the results with our previous analysis of Pol II stalling at a DNA photolesion, a TT cyclobutane pyrimidine dimer 8 (CPD, Fig. 1a), reveals that the two types of dinucleotide lesions trigger transcriptional stalling by different mechanisms. RESULTS Structure of a cisplatin-damaged Pol II elongation complexTo elucidate recognition of cisplatin-induced DNA damage by transcribing Pol II, we carried out a structure-function analysis of elongation complexes containing a cisplatin lesion in the template DNA strand. Elongation complexes were reconstituted from the 12-subunit S. cerevisiae Pol II and nucleic acid scaffolds as described 8,9 .A cisplatin lesion was first incorporated at registers +2/+3 of the template strand, directly downstream of the NTP-binding site at register +1 (scaffold A, Fig. 1b). The crystal structure of the resulting cisplatin-damaged elongation complex (complex A) was determined at 3.8-Å resolution (Fig. 1c,d and Methods). A very strong peak in the anomalous differe...

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Section: Discussionmentioning

confidence: 99%

Mechanism of transcriptional stalling at cisplatin-damaged DNA

Damsma

Alt

Brueckner

et al. 2007

Nat Struct Mol Biol

160

133

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“…As summarized in Figure 2, CSB interacts with several proteins involved in TCR/NER pathway. Recent studies also indicate that TCR can be accomplished without dissociation of RNAPII, and in this model CSB might help remodeling the RNAPII complex without release, allowing repair and the resumption of transcription (Laine et al, 2006a;Laine et al, 2006b;Tremeau-Bravard et al, 2004).…”

Section: The Role Of Csb In Various Cellular Processesmentioning

confidence: 99%

The role of Cockayne Syndrome group B (CSB) protein in base excision repair and aging

Stevnsner

Müftüoğlu

Aamann

et al. 2008

Mechanisms of Ageing and Development

126

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Cockayne Syndrome (CS) is a rare human genetic disorder characterized by progressive multisystem degeneration and segmental premature aging. The CS complementation group B (CSB) protein is engaged in transcription coupled and global nucleotide excision repair, base excision repair and general transcription. However, the precise molecular function of the CSB protein is still unclear. In the current review we discuss the involvement of CSB in some of these processes, with focus on the role of CSB in repair of oxidative damage, as deficiencies in the repair of these lesions may be an important aspect of the premature aging phenotype of CS.

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“…To prove or disprove this hypothesis, in vitro experiments have been performed; however the results are somewhat conflicting as they revealed either an inhibitory effect or no effect of blocked transcription on repair or dual incision [25,36,70]. In vitro studies by Tremeau -Bravard et al [70] and Laine and Egly [71] also indicate that TC-NER can be carried out without dissociation of the RNAPII (also reviewed in Laine and Egly [72]). However it is conceivable that TC-NER without displacement of RNAPIIo somehow requires conformational changes of the RNA polymerase to allow access to the DNA lesion and to resume transcription.…”

Section: Dna Damage and Stalled Rnapiiomentioning

confidence: 99%

“…As mentioned above, several studies suggest that the mammalian TC-NER complex is built up without displacement of RNAPIIo [41,70,71,77]. Importantly, the stalled RNAPIIo at the lesion may not be a sufficient barrier to prevent the DNA lesion from being accessible to repair proteins [54,71].…”

Section: Transcription Restart: a Crucial Role For Tfiis?mentioning

confidence: 99%

Transcription-coupled nucleotide excision repair in mammalian cells: molecular mechanisms and biological effects

2008

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The encounter of elongating RNA polymerase II (RNAPIIo) with DNA lesions has severe consequences for the cell as this event provides a strong signal for P53-dependent apoptosis and cell cycle arrest. To counteract prolonged blockage of transcription, the cell removes the RNAPIIo-blocking DNA lesions by transcription-coupled repair (TC-NER), a specialized subpathway of nucleotide excision repair (NER). Exposure of mice to UVB light or chemicals has elucidated that TC-NER is a critical survival pathway protecting against acute toxic and long-term effects (cancer) of genotoxic exposure. Deficiency in TC-NER is associated with mutations in the CSA and CSB genes giving rise to the rare human disorder Cockayne syndrome (CS). Recent data suggest that CSA and CSB play differential roles in mammalian TC-NER: CSB as a repair coupling factor to attract NER proteins, chromatin remodellers and the CSA-E3-ubiquitin ligase complex to the stalled RNAPIIo. CSA is dispensable for attraction of NER proteins, yet in cooperation with CSB is required to recruit XAB2, the nucleosomal binding protein HMGN1 and TFIIS. The emerging picture of TC-NER is complex: repair of transcription-blocking lesions occurs without displacement of the DNA damage-stalled RNAPIIo, and requires at least two essential assembly factors (CSA and CSB), the core NER factors (except for XPC-RAD23B), and TC-NER specific factors. These and yet unidentified proteins will accomplish not only efficient repair of transcription-blocking lesions, but are also likely to contribute to DNA damage signalling events.

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Fate of RNA Polymerase II Stalled at a Cisplatin Lesion

Cited by 63 publications

References 40 publications

Mechanism of transcriptional stalling at cisplatin-damaged DNA

Mechanism of transcriptional stalling at cisplatin-damaged DNA

The role of Cockayne Syndrome group B (CSB) protein in base excision repair and aging

Transcription-coupled nucleotide excision repair in mammalian cells: molecular mechanisms and biological effects

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