Binding of FANCI-FANCD2 Complex to RNA and R-Loops Stimulates Robust FANCD2 Monoubiquitination

Liang, Zhuobin; Liang, Fushun; Teng, Yaqun; Chen, Xiaoyong; Liu, Jingchun; Longerich, Simonne; Rao, Timsi; Green, Allison; Collins, Natalie B.; Xiong, Yong; Lan, Li; Sung, Patrick; Kupfer, Gary M.

doi:10.1016/j.celrep.2018.12.084

Cited by 69 publications

(70 citation statements)

References 36 publications

(61 reference statements)

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“…KR generates reactive oxygen species (ROS) through the excited chromophore and induces DNA damage and transcriptional activation at the genome‐integrated tet response element (TRE) locus in the U2OS TRE cells. Elevated R‐loop at the TRE locus over background is visualized by S9.6 immunofluorescence (Teng et al , ; Liang et al , ). DDX5 or PRMT5 knockdown (Fig A) led to a significant increase in R‐loop specifically at the TA‐KR marked locus, while the level of R‐loops was similar to the control at the TetR‐KR locus (Fig B and C).…”

Section: Resultsmentioning

confidence: 99%

Arginine methylation of the DDX 5 helicase RGG / RG motif by PRMT 5 regulates resolution of RNA:DNA hybrids

et al. 2019

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Aberrant transcription‐associated RNA : DNA hybrid (R‐loop) formation often causes catastrophic conflicts during replication, resulting in DNA double‐strand breaks and genomic instability. Preventing such conflicts requires hybrid dissolution by helicases and/or RN ase H. Little is known about how such helicases are regulated. Herein, we identify DDX 5, an RGG / RG motif‐containing DEAD ‐box family RNA helicase, as crucial player in R‐loop resolution. In vitro , recombinant DDX 5 resolves R‐loops in an ATP ‐dependent manner, leading to R‐loop degradation by the XRN 2 exoribonuclease. DDX 5‐deficient cells accumulate R‐loops at loci with propensity to form such structures based on RNA : DNA immunoprecipitation ( DRIP )‐ qPCR , causing spontaneous DNA double‐strand breaks and hypersensitivity to replication stress. DDX 5 associates with XRN 2 and resolves R‐loops at transcriptional termination regions downstream of poly(A) sites, to facilitate RNA polymerase II release associated with transcriptional termination. Protein arginine methyltransferase 5 ( PRMT 5) binds and methylates DDX 5 at its RGG / RG motif. This motif is required for DDX 5 interaction with XRN 2 and repression of cellular R‐loops, but not essential for DDX 5 helicase enzymatic activity. PRMT 5‐deficient cells accumulate R‐loops, resulting in increased formation of γH2 AX foci. Our findings exemplify a mechanism by which an RNA helicase is modulated by arginine methylation to resolve R‐loops, and its potential role in regulating transcription.

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Section: Resultsmentioning

confidence: 99%

Arginine methylation of the DDX 5 helicase RGG / RG motif by PRMT 5 regulates resolution of RNA:DNA hybrids

et al. 2019

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“…Therefore, we propose that CFS loci behave as both cis- and trans-acting components of the UPR that become unstable above a threshold of UPR activation and replication stress. The encounters between replication and transcription and R-loop formation would generate the substrate for FANCD2 binding and/or retention at CFSs and trigger the activation of the FANC pathway 18, 23, 24, 60 , constituting a metabolic and genome surveillance checkpoint.…”

Section: Discussionmentioning

confidence: 99%

FANCD2 tunes the UPR preventing mitochondrial stress-induced common fragile site instability

Fernandes

Miotto

Saint‐Ruf

et al. 2019

Preprint

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38Common fragile sites (CFSs) are genomic regions frequently involved in cancer-associated 39 rearrangements. Most CFSs lie within large genes, and their instability relies on 40 transcription-and replication-dependent mechanisms. Here, we uncover a role for the 41 UBL5-dependent branch of the unfolded protein response pathway (UPR) in the 42 maintenance of CFS stability. We show that genetic or pharmacological UPR activation 43 induces CFS gene expression and concomitant relocalization of FANCD2, a master 44 regulator of CFS stability, to CFSs. Furthermore, a genomic analysis of FANCD2 binding 45 sites identified an enrichment for mitochondrial UPR transcriptional response elements in 46 FANCD2 bound regions. We demonstrated that depletion of FANCD2 increases CFS gene 47 transcription and their instability while also inducing mitochondrial dysfunction and triggering 48 the activation of the UPR pathway. Depletion of UBL5, a mediator of the UPR, but not ATF4, 49reduces CFS gene expression and breakage in FANCD2-depleted cells. We thus 50 demonstrate that FANCD2 recruitment and function at CFSs depends on transcription and 51 UPR signaling, and in absence of transcription or UBL5, FANCD2 is dispensable for CFS 52 stability. We propose that FANCD2 coordinates nuclear and mitochondrial activities by 53 tuning the UPR to prevent genome instability. 54 55 56 57 58 CFS gene transcription and rescues chromosome fragility. Conversely, genetic or 127 pharmacological activation of the UPR induces CFS gene transcription and FANCD2 128 relocalization to CFS genes, dampening the UPR and preventing CFS instability. FANCD2 129 binding to CFS is dependent on CFS gene transcription and increases in a dose-dependent 130 manner. In addition, we show that FANCD2 is dispensable for maintaining CFS stability in 131 the absence of transcription. Mechanistically, we demonstrate that the UPR-dependent 132 induction of CFS genes is mediated by Ubiquitin-Like Protein 5 (UBL5), which was 133 previously shown to be involved in mitochondrial UPR signaling in C. elegans, and that 134 breaking up this signaling partially restores chromosome stability. We propose that CFSs 135 are part of a metabolic checkpoint, and by tuning the UPR with CFS replication, FANCD2 136 promotes metabolic homeostasis and genome integrity. 137 138 Results 140mRNA expression ATF4

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“…ALS/FTD, Fragile X syndrome, and Friedreich's Ataxia) (58)(59)(60)(61). Such tracts are known to induce DNA damage and mutations, as they are often stabilized upon the knock-down of certain factors involved in the transcription-coupled nucleotide excision repair (48), homologous recombination (62), and the Fanconi Anemia pathway (53,63). It is also known that multiple surveillance mechanisms in eukaryotes exist to eliminate the buildup of R-loops and the consequent DNA hyper-recombination (64).…”

Section: Discussionmentioning

confidence: 99%

Regulation of transcription termination by FUS and TDP-43

Zhao

et al. 2019

Preprint

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The carboxy-terminal domain (CTD) of the RNA polymerase II (RNAPII) subunit POLR2A is a platform for modifications specifying the recruitment of factors that regulate transcription, mRNA processing, and chromatin remodelling. We previously found that symmetrical dimethylation (me2s) of a CTD Arginine residue (R1810 in human) causes recruitment of the Tudor domain of SMN, which interacts with Senataxin. SMN is mutated in spinal muscular atrophy (SMA), and Senataxin is sometimes mutated in Amyotrophic Lateral Sclerosis (ALS). R1810me2s and SMN, like Senataxin, are important for resolving R-loops (DNA:RNA hybrids) at transcription terminators. FUS and TDP-43 (TARDBP) are DNA/RNA binding proteins that are sometimes mutated in ALS and FTD (Frontotemporal dementia). Here we show that TDP-43 and, to some extent, FUS are recruited by the R1810me2s-SMN pathway. Defects in FUS and TDP-43 recruitment influence RNAPII termination and R-loop accumulation, leading to elevated DNA damage at terminators that may contribute to neurodegenerative disorders like ALS and FTD.

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Binding of FANCI-FANCD2 Complex to RNA and R-Loops Stimulates Robust FANCD2 Monoubiquitination

Cited by 69 publications

References 36 publications

Arginine methylation of the DDX 5 helicase RGG / RG motif by PRMT 5 regulates resolution of RNA:DNA hybrids

Arginine methylation of the DDX 5 helicase RGG / RG motif by PRMT 5 regulates resolution of RNA:DNA hybrids

FANCD2 tunes the UPR preventing mitochondrial stress-induced common fragile site instability

Regulation of transcription termination by FUS and TDP-43

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Binding of FANCI-FANCD2 Complex to RNA and R-Loops Stimulates Robust FANCD2 Monoubiquitination

Cited by 69 publications

References 36 publications

Arginine methylation of the DDX 5 helicase RGG / RG motif by PRMT 5 regulates resolution of RNA:DNA hybrids

Arginine methylation of the DDX 5 helicase RGG / RG motif by PRMT 5 regulates resolution of RNA:DNA hybrids

FANCD2 tunes the UPR preventing mitochondrial stress-­induced common fragile site instability

Regulation of transcription termination by FUS and TDP-43

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FANCD2 tunes the UPR preventing mitochondrial stress-induced common fragile site instability