Search

Nucleotide excision repair (NER) counteracts the onset of cancer and aging by removing helix-distorting DNA lesions via a "cut-and-patch"-type reaction. The regulatory mechanisms that drive NER through its successive damage recognition, verification, incision, and gap restoration reaction steps remain elusive. Here, we show that the RAD5-related translocase HLTF facilitates repair through active eviction of incised damaged DNA together with associated repair proteins. Our data show a dual-incision-dependent recruitment of HLTF to the NER incision complex, which is mediated by HLTF's HIRAN domain that binds 3'-OH single-stranded DNA ends. HLTF's translocase motor subsequently promotes the dissociation of the stably damage-bound incision complex together with the incised oligonucleotide, allowing for an efficient PCNA loading and initiation of repair synthesis. Our findings uncover HLTF as an important NER factor that actively evicts DNA damage, thereby providing additional quality control by coordinating the transition between the excision and DNA synthesis steps to safeguard genome integrity., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Villafáfila lakes are a natural reserve included in the intergovernmental RAMSAR agreements for conservation of wetlands, with special interest for their brackish-saline waters. These lakes are located at the western margin of the Duero basin, whose aquifer system has no evaporitic rocks upstream. Understanding the origin of the lake's salinity, the groundwater circulation and the distribution of the brackish-saline waters in the area is important not only for the preservation and management of the natural reserve, but for human water consumption as well. Three types of waters have been identified according to their chemical composition. Type 1 are calcium-bicarbonate fresh waters identified in the local recharge areas (surrounding hills); Type 2 are mixed waters dominated by sodium and chloride-bicarbonate, identified at the toe of the hills; Type 3 are brackish to saline sodium-chloride waters from the lakes, springs and boreholes. Time domain electromagnetic (TDEM) profiles have revealed the existence of a basement elevation that forces brackish regional groundwater flow to rise. Radiocarbon age of regional groundwaters points to residence times of 20-30 Ky. Villafáfila lakes are through-flow lakes nourished by meteoric waters (direct precipitation and shallow groundwaters) as deduced by stable isotopes (δ 18 O H2O , δD H2O ), while the solutes are provided by ascendant deep groundwater flows in the lakes bottom and in the surrounding area. Sulphate stable isotopes (δ 18 O SO4= ; δ 34 S SO4= ) suggest that deep groundwaters have been in contact with Triassic and Cenozoic evaporites. Below the lake's bottom there is a brine (TDS = 27 g/L) contained within the lake-sediment aquitard that is concentrated by evaporation in the vadose zone and by salt recycling. A salinity inversion has been observed below the brine. The lack of saline crusts on the lake's bottom is favored by the SW outflow of the brine., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

THO/TREX is a conserved complex with a role in messenger ribonucleoprotein biogenesis that links gene expression and genome instability. Here, we show that human THO interacts with MFAP1 (microfibrillar-associated protein 1), a spliceosome-associated factor. Interestingly, MFAP1 depletion impairs cell proliferation and genome integrity, increasing γH2AX foci and DNA breaks. This phenotype is not dependent on either transcription or RNA-DNA hybrids. Mutations in the yeast orthologous gene SPP381 cause similar transcription-independent genome instability, supporting a conserved role. MFAP1 depletion has a wide effect on splicing and gene expression in human cells, determined by transcriptome analyses. MFAP1 depletion affects a number of DNA damage response (DDR) genes, which supports an indirect role of MFAP1 on genome integrity. Our work defines a functional interaction between THO and RNA processing and argues that splicing factors may contribute to genome integrity indirectly by regulating the expression of DDR genes rather than by a direct role., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Different proteins associate with the nascent RNA and the RNA polymerase (RNAP) to catalyze the transcription cycle and RNA export. If these processes are not properly controlled, the nascent RNA can thread back and hybridize to the DNA template forming R-loops capable of stalling replication, leading to DNA breaks. Given the transcriptional promiscuity of the genome, which leads to large amounts of RNAs from mRNAs to different types of ncRNAs, these can become a major threat to genome integrity if they form R-loops. Consequently, cells have evolved nuclear factors to prevent this phenomenon that includes THO, a conserved eukaryotic complex acting in transcription elongation and RNA processing and export that upon inactivation causes genome instability linked to R-loop accumulation. We revise and discuss here the biological relevance of THO and a number of RNA helicases, including the THO partner UAP56/DDX39B, as a paradigm of the cellular mechanisms of cotranscriptional R-loop prevention., (© 2019 Luna et al.; Published by Cold Spring Harbor Laboratory Press.)

R-loops, formed by co-transcriptional DNA-RNA hybrids and a displaced DNA single strand (ssDNA), fulfill certain positive regulatory roles but are also a source of genomic instability. One key cellular mechanism to prevent R-loop accumulation centers on the conserved THO/TREX complex, an RNA-binding factor involved in transcription elongation and RNA export that contributes to messenger ribonucleoprotein (mRNP) assembly, but whose precise function is still unclear. To understand how THO restrains harmful R-loops, we searched for new THO-interacting factors. We found that human THO interacts with the Sin3A histone deacetylase complex to suppress co-transcriptional R-loops, DNA damage, and replication impairment. Functional analyses show that histone hypo-acetylation prevents accumulation of harmful R-loops and RNA-mediated genomic instability. Diminished histone deacetylase activity in THO- and Sin3A-depleted cell lines correlates with increased R-loop formation, genomic instability, and replication fork stalling. Our study thus uncovers physical and functional crosstalk between RNA-binding factors and chromatin modifiers with a major role in preventing R-loop formation and RNA-mediated genome instability., (© 2017 The Authors.)