Senataxin, defective in ataxia oculomotor apraxia type 2, is involved in the defense against oxidative DNA damage

Suraweera, Amila; Bécherel, Olivier J.; Chen, Philip; Rundle, Natalie T.; Woods, Rick; Nakamura, Jun; Gatei, Magtouf; Criscuolo, Chiara; Filla, Alessandro; Chessa, Luciana; Fusser, Markus; Epe, Bernd; Gueven, Nuri; Lavin, Martin F.

doi:10.1083/jcb.200701042

Cited by 175 publications

(171 citation statements)

References 65 publications

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“…DDX1 did not coimmunoprecipitate with senataxin after IR treatment (see Fig. S8B), in agreement with the finding that senataxin does not contribute to cell survival post-IR (55).…”

Section: Ddx1 Contributes To Dsb Repair and Cell Survival Post-irsupporting

confidence: 78%

“…We therefore examined whether DDX1 colocalizes with two known markers associated with R-loops: the S9.6 antibody, which specifically recognizes RNA-DNA hybrids (53), and senataxin, an RNA helicase that resolves aberrant R-loops at transcriptionally active chromatin to facilitate proper transcription termination (54). Consistent with previous reports (27,55), immunostaining with both the S9.6 and antisenataxin antibodies revealed a speckled nuclear pattern without distinct foci (see Fig. S7 and S8 in the supplemental material).…”

Section: Ddx1 Contributes To Dsb Repair and Cell Survival Post-irsupporting

confidence: 52%

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DEAD Box 1 Facilitates Removal of RNA and Homologous Recombination at DNA Double-Strand Breaks

Germain

Poon

et al. 2016

Molecular and Cellular Biology

123

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Although RNA and RNA-binding proteins have been linked to double-strand breaks (DSBs), little is known regarding their roles in the cellular response to DSBs and, if any, in the repair process. Here, we provide direct evidence for the presence of RNA-DNA hybrids at DSBs and suggest that binding of RNA to DNA at DSBs may impact repair efficiency. Our data indicate that the RNAunwinding protein DEAD box 1 (DDX1) is required for efficient DSB repair and cell survival after ionizing radiation (IR), with depletion of DDX1 resulting in reduced DSB repair by homologous recombination (HR). While DDX1 is not essential for end resection, a key step in homology-directed DSB repair, DDX1 is required for maintenance of the single-stranded DNA once generated by end resection. We show that transcription deregulation has a significant effect on DSB repair by HR in DDX1-depleted cells and that RNA-DNA duplexes are elevated at DSBs in DDX1-depleted cells. Based on our combined data, we propose a role for DDX1 in resolving RNA-DNA structures that accumulate at DSBs located at sites of active transcription. Our findings point to a previously uncharacterized requirement for clearing RNA at DSBs for efficient repair by HR. DEAD box proteins are a family of putative RNA helicases that function by altering RNA secondary structure. This protein family has been implicated in all aspects of RNA metabolism. The DEAD box 1 gene (DDX1) is a widely expressed gene that is misexpressed in a number of cancers, including retinoblastoma, neuroblastoma, and breast cancer (1, 2). Knockout of DDX1 leads to early embryonic lethality in mice and severely reduced fertility in flies (3, 4). DDX1 is involved in the transport of RNAs from the nucleus to the cytoplasm and regulates cytoplasmic localization of the splicing-regulatory protein KSRP (5). In neurons, DDX1 resides in RNA-transporting granules, cytoplasmic organelles that regulate the localization and expression of target mRNAs (6, 7). DDX1 has also been identified as a core subunit of the human tRNA ligase complex which is essential for tRNA splicing (8).In addition to its roles in RNA metabolism, DDX1 has been implicated in the cellular response to DNA double-strand breaks (DSBs). Upon treatment of cells with ionizing radiation (IR), DDX1 rapidly accumulates at a subset of DNA DSBs (ϳ30%), where it forms IR-induced foci that colocalize with ␥-H2AX, a marker for DSBs (9). DDX1 coimmunoprecipitates with the MRN (MRE11-RAD50-NBS1) complex, the early sensor of DNA DSBs, and ATM (ataxia telangiectasia mutated) protein, the key transducer of the signaling cascade in response to DSBs (10, 11). DSBs induce DDX1 phosphorylation in an ATM-dependent manner. Notably, IR-induced DDX1 foci are lost when cells are treated with RNase H, an enzyme that specifically digests RNA from RNA-DNA hybrids (9). These results suggest that RNA-DNA double-stranded structures are required for the presence and/or retention of DDX1 at DSBs. In line with this observation, biochemical analysis has shown that DDX1 can unwind both...

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“…DDX1 did not coimmunoprecipitate with senataxin after IR treatment (see Fig. S8B), in agreement with the finding that senataxin does not contribute to cell survival post-IR (55).…”

Section: Ddx1 Contributes To Dsb Repair and Cell Survival Post-irsupporting

confidence: 78%

Section: Ddx1 Contributes To Dsb Repair and Cell Survival Post-irsupporting

confidence: 52%

DEAD Box 1 Facilitates Removal of RNA and Homologous Recombination at DNA Double-Strand Breaks

Germain

Poon

et al. 2016

Molecular and Cellular Biology

123

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“…Interestingly, mutations in SETX lead to two neurological disorders, an autosomal dominant form, ALS4 (amyotrophic lateral sclerosis type 4) (Blair et al 2000), and an autosomal recessive form, AOA2 (ataxiaoculomotor apraxia type 2) (Moreira et al 2004;Chen et al 2006). Although the only characterized role of SETX in humans is its involvement in DNA damage repair (Suraweera et al 2007), it will be interesting to determine whether SETX also functions in transcription termination, particularly in neurons.…”

Section: Rat1/xrn2: the Transcription Termination Torpedomentioning

confidence: 99%

Transcription termination by nuclear RNA polymerases

Richard

Manley²

2009

Genes Dev.

285

325

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Gene transcription in the cell nucleus is a complex and highly regulated process. Transcription in eukaryotes requires three distinct RNA polymerases, each of which employs its own mechanisms for initiation, elongation, and termination. Termination mechanisms vary considerably, ranging from relatively simple to exceptionally complex. In this review, we describe the present state of knowledge on how each of the three RNA polymerases terminates and how mechanisms are conserved, or vary, from yeast to human.

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“…Mutations in human SETX (senataxin), the ortholog of yeast SEN1, cause two clinically distinct neurological diseases, ataxia-ocular apraxia 2 and juvenile amyotrophic lateral sclerosis (Chen et al 2004(Chen et al , 2006Moreira et al 2004;Duquette et al 2005;Suraweera et al 2007;Suraweera et al 2009). The yeast and human proteins are strikingly similar in their organization.…”

mentioning

confidence: 99%

Sen1p Performs Two Genetically Separable Functions in Transcription and Processing of U5 Small Nuclear RNA inSaccharomyces cerevisiae

et al. 2010

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The Saccharomyces cerevisiae SEN1 gene codes for a nuclear-localized superfamily I helicase. SEN1 is an ortholog of human SETX (senataxin), which has been implicated in the neurological disorders ataxiaocular apraxia type 2 and juvenile amyotrophic lateral sclerosis. Pleiotropic phenotypes conferred by sen1 mutations suggest that Sen1p affects multiple steps in gene expression. Sen1p is embedded in a proteinprotein interaction network involving direct binding to multiple partners. To test whether the interactions occur independently or in a dependent sequence, we examined interactions with the RNA polymerase II subunit Rpb1p, which is required for transcription, and Rnt1p, which is required for 39-end maturation of many noncoding RNAs. Mutations were identified that impair one of the two interactions without impairing the other interaction. The effects of the mutants on the synthesis of U5 small nuclear RNA were analyzed. Two defects were observed, one in transcription termination and one in 39-end maturation. Impairment of the Sen1p-Rpb1p interaction resulted in a termination defect. Impairment of the Sen1p-Rnt1p interaction resulted in a processing defect. The results suggest that the Sen1p-Rpb1p and Sen1p-Rnt1p interactions occur independently of each other and serve genetically separable purposes in targeting Sen1p to function in two temporally overlapping steps in gene expression.

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Senataxin, defective in ataxia oculomotor apraxia type 2, is involved in the defense against oxidative DNA damage

Cited by 175 publications

References 65 publications

DEAD Box 1 Facilitates Removal of RNA and Homologous Recombination at DNA Double-Strand Breaks

DEAD Box 1 Facilitates Removal of RNA and Homologous Recombination at DNA Double-Strand Breaks

Transcription termination by nuclear RNA polymerases

Sen1p Performs Two Genetically Separable Functions in Transcription and Processing of U5 Small Nuclear RNA inSaccharomyces cerevisiae

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