Biology subjects

Gonzalez-Vinceiro, L.

Publications and source records attributed to Gonzalez-Vinceiro, L..

3 recordsLinked to original sources

bioRxiv2026

RNF25 Ubiquitin E3 activity Safeguards Genome Integrity by Modulating DNA Replication, Transcription and Translation.

The human genome encodes several hundreds of ubiquitin E3 enzymes, most of which have poorly understood biological roles. In search for novel ubiquitin E3 enzymes involved in DNA damage tolerance, we identified RNF25 as a candidate to have a role in DNA replication stress tolerance. Under stress conditions, RNF25 translocates to the nucleus in a cGAS-dependent manner, and loss of RNF25 ubiquitin E3 activity leads to the accumulation of replication-dependent ssDNA gaps. Combining functional assays with with mass-spectrometry based proteomics approaches such as TULIP2, iPOND-MS and TurboID, we found that, mechanistically, RNF25 promotes the stability of the replication fork at Transcription-Replication Conflicts by the ubiquitin-mediated clearance of RAD18, mono-ubiquitinated PCNA, H2B-K120ub and RECQL, among others. Lack of RNF25 ubiquitin E3 activity promotes the occurrence of Transcription-Replication Conflicts and destabilizes reversed replication forks, enabling re-priming and accumulation of ssDNA gaps behind the replication fork, ultimately compromising genome integrity. Overall, RNF25 regulates DNA replication, transcription and translation in an ubiquitin-based goldilocks.

molecular biology↗

bioRxiv2025

miRNA-mediated cell-to-cell communications boost DNA repair during theRadioadaptative Response

The Radioadaptive Response (RAR) is a phenomenon where a low, or priming, dose of ionizing radiation enhances cellular resistance to subsequent higher doses. We investigated whether RAR involves alterations in Homologous Recombination (HR), a high-fidelity DNA repair pathway. Using fibroblast models, we found that primed cells exhibit accelerated DNA end resection, an initial and essential HR step. This effect is mostly mediated by a bystander mechanism involving small extracellular vesicles (sEVs), as conditioned media fully replicated it. RNA profiling of sEVs identified miR-126-3p and miR-451a as key regulators of this response. Significantly, inhibiting miR-451a induced RAR in normally unresponsive cells. We further identified a miR-451a-p38-CCAR2 axis that enhances HR through suppression of CCAR2. These findings delineate a novel miRNA-mediated, sEV-driven mechanism that regulates HR during RAR, with potential therapeutic implications. Significance StatementWe uncover a previously unrecognized mechanism by which human fibroblasts enhance DNA double-strand break repair through homologous recombination following a priming dose of ionizing radiation-a phenomenon known as the radioadaptive response (RAR). We demonstrate that this enhanced repair capacity is driven by small extracellular vesicle (sEV)-mediated intercellular communication, through a transient reprograming of the DNA repair capacity of neighboring cells by modulating the levels of two key microRNAs. These provide new clues on how extracellular RNA signaling governs genome maintenance, with significant implications for genome stability in healthy and pathological context. The identification of actionable modulators further strengthens the translational potential of our work.

molecular biology↗

bioRxiv2025

PLAMseq enables the proteo-genomic characterization of chromatin-associated proteins and protein interactions in a single experimental workflow.

Chromatin Immunoprecipitation (ChIP) and Co-Immunoprecipitation (CoIP) assays are common approaches to characterize the genomic localization and protein interactors, respectively, for a protein of interest. However, these approaches require the use of specific antibodies, which often face sensitivity and specificity issues. Based on TurboID, we developed PLAMseq (Proximity Labelled Affinity-purified Mass spectrometry plus sequencing), which enables, in the same workflow, to identify the genomic loci and the interacting proteome of a protein of interest. Moreover, PLAMseq can also be applied to specifically map protein interactions and ubiquitin(-like) modified proteins. We validated PLAMseq with two well characterized proteins, RNA polymerase II and CTCF, with excellent robustness and reproducibility. Next, we applied PLAMseq to characterize Histone H1 SUMOylation, which study has remained elusive due to the lack of specific reagents, and found that SETDB1 binds to SUMOylated histone H1.2 and H1.4 which also colocalize with H3K9me3 at repetitive regions of the genome.

molecular biology↗