2025 Hyper Recent •CC0 1.0 Universal

This work is dedicated to the public domain. No rights reserved.

Access Preprint From Server
July 1st, 2025
Version: 2
Ben Gurion University of the Negev
molecular biology
biorxiv

RAD18 Methylation by the Methyltransferase SETD6 Attenuates DNA Breaks

Weil, L. E.Open in Google Scholar•Feldman, M.Open in Google Scholar•Duine, J. V.Open in Google Scholar•Qiu, J.Open in Google Scholar•LaBaer, J.Open in Google Scholar•Levy, D.Open in Google Scholar

This study investigated the interaction between the SETD6 lysine methyltransferase and RAD18, a key protein in the DNA damage repair pathway. SETD6 belongs to the SET-domain-containing family of proteins, which are known to catalyze protein methylation, a post-translational modification that plays a critical role in regulating protein function, stability, and interactions. Using protein microarray technology, we identified RAD18 as an interactor and substrate of SETD6. We confirmed this interaction through ELISA and immunoprecipitation assays, demonstrating that SETD6 directly binds and methylates RAD18. Using mass spectrometry and site-directed mutagenesis, we identified that RAD18 undergoes mono-methylation at the K73 and K406 residues. Furthermore, we found that RAD18 methylation affects its nuclear localization. Specifically, SETD6 KO cells exhibited increased nuclear RAD18 levels, suggesting that methylation status influences RAD18s shuttling between the cytoplasm and nucleus. Notably, depletion of SETD6 led to elevated markers of DNA damage ({gamma}H2AX) and increased DNA breaks, as evidenced by comet assays. Restoring SETD6 activity significantly reduced DNA damage, while a catalytic inactive mutant did not have this effect, underscoring the importance of SETD6's enzymatic function. Overall, our results demonstrate that SETD6-mediated methylation of RAD18 is essential for attenuating DNA breaks, thereby regulating its cellular localization and function in maintaining genomic integrity.

Similar Papers

biorxiv
Wed Jul 02 2025
ChAHP Silences SINE Retrotransposons by Inhibiting TFIIIB Recruitment
Short interspersed nuclear elements (SINEs) are abundant non-autonomous transposable elements derived from RNA polymerase III (POL III)-transcribed short non-coding RNAs. SINEs retain sequence features recognized by the POL III machinery and constitute a substantial portion of vertebrate genomes. Despite their impact on genome stability and evolution, the mechanisms governing SINE transcription re...
Schnabl-Baumgartner, J.
•
Mohn, F.
•
Schwaiger, M.
•
Ahel, J.
...•
Buhler, M.
biorxiv
Wed Jul 02 2025
Natural Product-Like Fragments Unlock Novel Chemotypes for a Kinase Target - Exploring Options beyond the Flatland
In this study we utilized a high-performance soaking system of protein kinase A (PKA) to perform a crystallographic screening of a natural product-like fragment library. We resolved 36 fragmentbound structures, corresponding to a hit rate of 41%. Nine fragments bound within the ATP site, nine peripherally, and 18 interacted with both the ATP and peripheral sites. One fragment binds to the same sit...
Santura, A.
•
Mueller, J.
•
Wolter, M.
•
Tutzschky, I.-C.
...•
Czodrowski, P.
biorxiv
Wed Jul 02 2025
Latrophilin mediates flow activation of Notch to control vascular endothelial phenotype and atherosclerosis
Fluid shear stress (FSS) is a major determinant of endothelial cell (EC) phenotype, with physiological laminar FSS promoting arterial identity and stability, whereas disturbed FSS promotes atherosclerosis. We previously identified the adhesion G protein-coupled receptor (GPCR) Latrophilin-2 (Lphn2) as a junctional protein required for FSS activation of the PECAM1/VE-cadherin/VEGFR2/PlexinD1 juncti...
Tanaka, K.
•
Chen, M.
•
Chanduri, M. V.
•
Zhang, H.
...•
Schwartz, M. A.
biorxiv
Wed Jul 02 2025
SETD6 Mediates Selective Interaction and Genomic Occupancy of BRD4 and MITF in Melanoma cells
Aberrant transcriptional programs mediate malignant transformation of melanoma, the most aggressive form of skin cancer. The lysine methyltransferase SETD6 has been implicated in regulating transcription, cell adhesion, migration, and other processes in various cancers, however its role in melanoma remains unexplored. We recently reported that SETD6 mono-methylates the BRD4 at K99 to selectively r...
Biton, T. E.
•
Feldman, M.
•
Davidy, T.
•
Moskovitz, N. T.
...•
Levy, D.
biorxiv
Wed Jul 02 2025
Multiplexed single-cell transcriptomics reveals diverse phenotypic outcomes for pathogenic SHP2 variants
The protein tyrosine phosphatase SHP2, encoded by PTPN11, is an important regulator of Ras/MAPK signaling that acts downstream of receptor tyrosine kinases and other transmembrane receptors. Germline PTPN11 mutations cause developmental disorders such as Noonan Syndrome, whereas somatic mutations drive various cancers. While many pathogenic mutations enhance SHP2 catalytic activity, others are ina...
van Vlimmeren, A. E.
•
Giglio, R. M.
•
Jiang, Z.
•
Lee, M.
...•
Shah, N. H.
biorxiv
Wed Jul 02 2025
SF3B1K700E rewires splicing of cell-cycle regulators
Pre-mRNA splicing plays a crucial role in maintaining cellular homeostasis, with strict regulation required for processes such as cell cycle progression. SF3B1, a core component of the spliceosome, has emerged as a key player in alternative splicing regulation and is frequently mutated in cancer. Among these mutations, SF3B1K700E disrupts normal splicing patterns and deregulates cell cycle control...
Baker, M.
•
Sharma, A.
•
Engal, E.
•
Petasny, M.
...•
Salton, M.
biorxiv
Tue Jul 01 2025
Side-by-side systematic characterization of novel FPs in budding yeast
Fluorescent proteins (FPs) have become indispensable for biological research. Since the discovery of the first FP, Aequorea victoria green fluorescent protein (avGFP), new fluorescent proteins are continuously being developed. To achieve optimal experimental results, selecting an FP based on specific characteristics -such as brightness, photostability, photochromicity, monomericity, pH robustness,...
Botman, D.
•
de Groot, D. H.
•
Wehrens, M.
•
van Rossum, K.
...•
Teusink, B.
biorxiv
Tue Jul 01 2025
Plagl1 and Lrrc58 control mammalian body size by triggering target-directed microRNA degradation of miR-322 and miR-503
Precise control of microRNA (miRNA) expression is critical during development. An important mechanism of miRNA regulation is target-directed microRNA degradation (TDMD), a pathway in which the binding of miRNAs to specialized trigger RNAs induces ubiquitylation and decay of associated Argonaute (AGO) proteins by the ZSWIM8 ubiquitin ligase. Concomitant release of miRNAs results in their rapid turn...
LaVigne, C. A.
•
Han, J.
•
Zhang, H.
•
Cho, S.
...•
Mendell, J. T.
biorxiv
Tue Jul 01 2025
Multifaceted mRNA analysis using programmed RNA cleavage byMucilaginibacter paludis Argonaute
Argonaute (Ago) endonucleases are ubiquitous across all domains of life and, like Cas nucleases, can be programmed by guide oligonucleotides to target and cleave specific nucleic acid sequences. However, unlike Cas enzymes, Agos are not restricted by sequence requirements such as protospacer adjacent motifs (PAMs) enhancing their utility as molecular tools. This study demonstrates diverse applicat...
Liu, L.
•
Hunt, E.
•
Wolf, E. J.
•
Correa, I. R.
...•
Grunberg, S.