Biology subjects

Gregersen, L. H.

Publications and source records attributed to Gregersen, L. H..

3 recordsLinked to original sources

bioRxiv2025

SCAF1 driven polyadenylation site usage regulates mRNA isoform expression and neuronal differentiation

Accurate co-transcriptional processing is required for correct gene expression of mRNA transcript isoforms under unperturbed conditions, but particularly during development, to ensure tissue-specific mRNA isoform expression. Here we show that the poorly studied SR-related CTD-associated factors SCAF1 protein regulates polyadenylation site usage towards the end of genes. SCAF1 interacts directly with the phosphorylated C-terminal domain (CTD) of RNA polymerase II (RNAPII), in a complex enriched with elongation and 3 end processing factors. While SCAF1 knockout in HEK293 cells is innocuous, it leads to a shift towards expression of shorter mRNA transcripts by co-transcriptional usage of early polyadenylation sites. SCAF1 deficiency induced via auxin-dependent degradation in neuron differentiating mouse embryonic stem cells (mESCs) results in neuronal commitment defects, mediated by altered mRNA isoform usage that impacts expression of key neuronal genes. These findings highlight the importance of mRNA isoform usage and underscores the key role for SCAF proteins in its regulation though polyadenylation site selection.

molecular biology↗

bioRxiv2025

Oxidative stress triggers RNAPII arrest through PARylation and DNA damage

UV or gamma irradiation, as well as certain chemicals, generate DNA damage that disrupts transcription through a variety of well-characterised mechanisms. In contrast, the transcriptional response to oxidative stress remains poorly understood. Here, we describe a rapid and widespread shutdown of transcription following oxidative DNA base damage. By monitoring RNAPII occupancy and elongation dynamics, we demonstrate that oxidative stress temporarily halts RNAPII pause release and arrests the progression of elongation complexes within the gene body. We present evidence that this occurs in a unique and transient manner, characterised by abrupt arrest of elongating RNAPII dead in its tracks, followed by rapid transcriptional recovery as DNA lesions are repaired. We find that the restriction of initiation and early elongation complexes is regulated by PARylation, whereas recovery of RNAPII arrested within the gene body requires DNA repair mediated by the base excision repair (BER) and single-strand break repair (SSBR) pathways.

molecular biology↗

bioRxiv2024

RNA Polymerase II subunits overexpressions induce genome instability and deregulate transcription

Independently of the pathways or circuits deregulated in cancer cells, these present altered transcription patterns, often also direct consequence of deregulation of transcription factors. In this sense, also the RNA Polymerase complexes responsible for transcription can be affected in cancers. We find that upregulations of RNA Polymerase II subunits, especially the largest ones, correlates with poor cancer patients outcome across a range of tumor types, presenting increased genome instability. Overexpressing the subunits RPB1, RPB3 and RPB4 in cells we find that these induce DNA damage. However, the mechanisms behind this increased genome instability are specific for each subunit, linked to the unique transcription alterations generated by the subunit overexpression. Importantly, we find significant overlap between the genes with more DNA damage in our cell line models and those more affected in cancers with subunit upregulation, indicating that upregulations could be responsible for some of the phenotypes present in these patients.

molecular biology↗