Skip to main content

Research Repository

Advanced Search

All Outputs (28)

Saccharin disrupts bacterial cell envelope stability and interferes with DNA replication dynamics (2025)
Journal Article
de Dios, R., Gadar, K., Proctor, C. R., Maslova, E., Han, J., Soliman, M. A. N., Krawiel, D., Dunbar, E. L., Singh, B., Peros, S., Killelea, T., Warnke, A.-L., Haugland, M. M., Bolt, E. L., Lentz, C. S., Rudolph, C. J., & McCarthy, R. R. (2025). Saccharin disrupts bacterial cell envelope stability and interferes with DNA replication dynamics. EMBO Molecular Medicine, https://doi.org/10.1038/s44321-025-00219-1

Saccharin has been part of the human diet for over 100 years, and there is a comprehensive body of evidence demonstrating that it can influence the gut microbiome, ultimately impacting human health. However, the precise mechanisms through which sacch... Read More about Saccharin disrupts bacterial cell envelope stability and interferes with DNA replication dynamics.

The human HELQ helicase and XRN2 exoribonuclease cooperate in R-loop resolution (2025)
Journal Article
Pan, J. M., Betts, H., Cubbon, A., Bolt, E. L., Soultanas, P., & He, L. (2025). The human HELQ helicase and XRN2 exoribonuclease cooperate in R-loop resolution. Open Biology, 15(2), https://doi.org/10.1098/rsob.240112

The human HELQ helicase is a superfamily 2, 3’-5’ helicase homologous to POLQ and RNA helicases of the Ski2-like subfamily. It is involved in diverse aspects of DNA repair and is an emerging prognosis biomarker and novel drug-target for cancer therap... Read More about The human HELQ helicase and XRN2 exoribonuclease cooperate in R-loop resolution.

Identification of a novel nuclease activity in human DDX49 helicase (2024)
Journal Article
Parkes, A. J., Anandavijayan, S., Lou-Hing, A., Downs, O., Killelea, T., Martin, L., Kapllanaj, F., & Bolt, E. L. (2024). Identification of a novel nuclease activity in human DDX49 helicase. Royal Society Open Science, 11(12), Article 241891. https://doi.org/10.1098/rsos.241891

Human DDX49 is an emerging target in cancer progression and retroviral diseases through its essential roles in nucleolar RNA processing. Here, we identify nuclease activity of human DDX49, which requires active site aspartate residues within a conser... Read More about Identification of a novel nuclease activity in human DDX49 helicase.

POLD3 as Controller of Replicative DNA Repair (2024)
Journal Article
Alli, N., Lou-Hing, A., Bolt, E. L., & He, L. (2024). POLD3 as Controller of Replicative DNA Repair. International Journal of Molecular Sciences, 25(22), Article 12417. https://doi.org/10.3390/ijms252212417

Multiple modes of DNA repair need DNA synthesis by DNA polymerase enzymes. The eukaryotic B-family DNA polymerase complexes delta (Polδ) and zeta (Polζ) help to repair DNA strand breaks when primed by homologous recombination or single-strand DNA ann... Read More about POLD3 as Controller of Replicative DNA Repair.

Escherichia coli DNA repair helicase Lhr is also a uracil-DNA glycosylase (2023)
Journal Article
Buckley, R. J., Lou-Hing, A., Hanson, K. M., Ahmed, N. R., Cooper, C. D., & Bolt, E. L. (2023). Escherichia coli DNA repair helicase Lhr is also a uracil-DNA glycosylase. Molecular Microbiology, 120(2), 298-306. https://doi.org/10.1111/mmi.15123

DNA glycosylases protect genetic fidelity during DNA replication by removing potentially mutagenic chemically damaged DNA bases. Bacterial Lhr proteins are well-characterized DNA repair helicases that are fused to additional 600–700 amino acids of un... Read More about Escherichia coli DNA repair helicase Lhr is also a uracil-DNA glycosylase.

Escherichia coli DNA repair helicase Lhr is also a uracil‐DNA glycosylase (2023)
Journal Article
Buckley, R. J., Lou‐Hing, A., Hanson, K. M., Ahmed, N. R., Cooper, C. D. O., & Bolt, E. L. (2023). Escherichia coli DNA repair helicase Lhr is also a uracil‐DNA glycosylase. Molecular Microbiology, 120(2), 298-306. https://doi.org/10.1111/mmi.15123

DNA glycosylases protect genetic fidelity during DNA replication by removing potentially mutagenic chemically damaged DNA bases. Bacterial Lhr proteins are well‐characterized DNA repair helicases that are fused to additional 600–700 amino acids of un... Read More about Escherichia coli DNA repair helicase Lhr is also a uracil‐DNA glycosylase.

Archaeal Hel308 suppresses recombination through a catalytic switch that controls DNA annealing (2023)
Journal Article
Lever, R., Simmons, E., Gamble-Milner, R., Buckley, R., Harrison, C., Parkes, A., Mitchell, L., Gausden, J., Škulj, S., Bertoša, B., Bolt, E. L., & Allers, T. (2023). Archaeal Hel308 suppresses recombination through a catalytic switch that controls DNA annealing. Nucleic Acids Research, 51(16), 8563-8574. https://doi.org/10.1093/nar/gkad572

Hel308 helicases promote genome stability in archaea and are conserved in metazoans, where they are known as HELQ. Their helicase mechanism is well characterised, but it is unclear how they specifically contribute to genome stability in archaea. We s... Read More about Archaeal Hel308 suppresses recombination through a catalytic switch that controls DNA annealing.

Cas1-Cas2 physically and functionally interacts with DnaK to modulate CRISPR Adaptation (2023)
Journal Article
Killelea, T., Dimude, J. U., He, L., Stewart, A. L., Kemm, F. E., Radovčí, M., Ivančić-Baće, I., Rudolph, C. J., & Bolt, E. L. (2023). Cas1-Cas2 physically and functionally interacts with DnaK to modulate CRISPR Adaptation. Nucleic Acids Research, 51(13), 6914-6926. https://doi.org/10.1093/nar/gkad473

Prokaryotic Cas1-Cas2 protein complexes generate adaptive immunity to mobile genetic elements (MGEs), by capture and integration of MGE DNA in to CRISPR sites. De novo immunity relies on naive adaptation-Cas1-Cas2 targeting of MGE DNA without the aid... Read More about Cas1-Cas2 physically and functionally interacts with DnaK to modulate CRISPR Adaptation.

CRISPR-Cas adaptation in Escherichia coli (2023)
Journal Article
Mitic, D., Bolt, E. L., & Ivancic-Bace, I. (2023). CRISPR-Cas adaptation in Escherichia coli. Bioscience Reports, 43(3), Article BSR20221198. https://doi.org/10.1042/BSR20221198

Prokaryotes use the adaptive immunity mediated via the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR associated (CRISPR-Cas) system for protection against invading elements such as phages and plasmids. The immunity is achieved... Read More about CRISPR-Cas adaptation in Escherichia coli.

Interaction of human HelQ with DNA polymerase delta halts DNA synthesis and stimulates DNA single-strand annealing (2023)
Journal Article
He, L., Lever, R., Cubbon, A., Tehseen, M., Jenkins, T., Nottingham, A. O., Horton, A., Betts, H., Fisher, M., Hamdan, S. M., Soultanas, P., & Bolt, E. L. (2023). Interaction of human HelQ with DNA polymerase delta halts DNA synthesis and stimulates DNA single-strand annealing. Nucleic Acids Research, 51(4), 1740-1749. https://doi.org/10.1093/nar/gkad032

DNA strand breaks are repaired by DNA synthesis from an exposed DNA end paired with a homologous DNA template. DNA polymerase delta (Pol δ) catalyses DNA synthesis in multiple eukaryotic DNA break repair pathways but triggers genome instability unles... Read More about Interaction of human HelQ with DNA polymerase delta halts DNA synthesis and stimulates DNA single-strand annealing.

A Tryptophan ‘Gate’ in the CRISPR-Cas3 Nuclease Controls ssDNA Entry into the Nuclease Site, That When Removed Results in Nuclease Hyperactivity (2021)
Journal Article
He, L., Matošević, Z. J., Mitić, D., Markulin, D., Killelea, T., Matković, M., Bertoša, B., Ivančić-Baće, I., & Bolt, E. L. (2021). A Tryptophan ‘Gate’ in the CRISPR-Cas3 Nuclease Controls ssDNA Entry into the Nuclease Site, That When Removed Results in Nuclease Hyperactivity. International Journal of Molecular Sciences, 22(6), Article 2848. https://doi.org/10.3390/ijms22062848

Cas3 is a ssDNA-targeting nuclease-helicase essential for class 1 prokaryotic CRISPR immunity systems, which has been utilized for genome editing in human cells. Cas3-DNA crystal structures show that ssDNA follows a pathway from helicase domains into... Read More about A Tryptophan ‘Gate’ in the CRISPR-Cas3 Nuclease Controls ssDNA Entry into the Nuclease Site, That When Removed Results in Nuclease Hyperactivity.

The HelQ human DNA repair helicase utilizes a PWI-like domain for DNA loading through interaction with RPA, triggering DNA unwinding by the HelQ helicase core (2021)
Journal Article
Jenkins, T., Northall, S. J., Ptchelkine, D., Lever, R., Cubbon, A., Betts, H., Taresco, V., Cooper, C. D. O., McHugh, P. J., Soultanas, P., & Bolt, E. L. (2021). The HelQ human DNA repair helicase utilizes a PWI-like domain for DNA loading through interaction with RPA, triggering DNA unwinding by the HelQ helicase core. NAR Cancer, 3(1), Article zcaa043. https://doi.org/10.1093/narcan/zcaa043

Genome instability is a characteristic enabling factor for carcinogenesis. HelQ helicase is a component of human DNA maintenance systems that prevent or reverse genome instability arising during DNA replication. Here, we provide details of the molecu... Read More about The HelQ human DNA repair helicase utilizes a PWI-like domain for DNA loading through interaction with RPA, triggering DNA unwinding by the HelQ helicase core.

Integration of diverse DNA substrates by a casposase can be targeted to R-loops in vitro by its fusion to Cas9 (2021)
Journal Article
Lau, C. H., & Bolt, E. L. (2021). Integration of diverse DNA substrates by a casposase can be targeted to R-loops in vitro by its fusion to Cas9. Bioscience Reports, 41(1), Article BSR20203595. https://doi.org/10.1042/bsr20203595

CRISPR systems build adaptive immunity against mobile genetic elements by DNA capture and integration catalysed by Cas1–Cas2 protein complexes. Recent studies suggested that CRISPR repeats and adaptation module originated from a novel type of DNA tra... Read More about Integration of diverse DNA substrates by a casposase can be targeted to R-loops in vitro by its fusion to Cas9.

Mechanistic insights into Lhr helicase function in DNA repair (2020)
Journal Article
Buckley, R., Kramm, K., Cooper, C. D. O., Grohmann, D., & Bolt, E. L. (2020). Mechanistic insights into Lhr helicase function in DNA repair. Biochemical Journal, 477(16), 2935–2947. https://doi.org/10.1042/bcj20200379

The DNA helicase Lhr is present throughout archaea, including in the Asgard and Nanoarchaea, and has homologues in bacteria and eukaryotes. It is thought to function in DNA repair but in a context that is not known. Our data show that archaeal Lhr pr... Read More about Mechanistic insights into Lhr helicase function in DNA repair.

A gold standard, CRISPR/Cas9-based complementation strategy reliant on 24 nucleotide bookmark sequences (2020)
Journal Article
Seys, F. M., Rowe, P., Bolt, E. L., Humphreys, C. M., & Minton, N. P. (2020). A gold standard, CRISPR/Cas9-based complementation strategy reliant on 24 nucleotide bookmark sequences. Genes, 11(4), Article 458. https://doi.org/10.3390/genes11040458

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. Phenotypic complementation of gene knockouts is an essential step in establishing function. Here, we describe a simple strategy for ‘gold standard’ complementation in which the mutant allele i... Read More about A gold standard, CRISPR/Cas9-based complementation strategy reliant on 24 nucleotide bookmark sequences.

Cas3 Protein—A Review of a Multi-Tasking Machine (2020)
Journal Article
He, L., St. John James, M., Radovcic, M., Ivancic-Bace, I., & Bolt, E. L. (2020). Cas3 Protein—A Review of a Multi-Tasking Machine. Genes, 11(2), Article 208. https://doi.org/10.3390/genes11020208

Cas3 has essential functions in CRISPR immunity but its other activities and roles, in vitro and in cells, are less widely known. We offer a concise review of the latest understanding and questions arising from studies of Cas3 mechanism during CRISPR... Read More about Cas3 Protein—A Review of a Multi-Tasking Machine.

Adaptation processes that build CRISPR immunity: creative destruction, updated (2019)
Journal Article
Lau, C., Reeves, R., & Bolt, E. (2019). Adaptation processes that build CRISPR immunity: creative destruction, updated. Essays in Biochemistry, 63(2), 227-235. https://doi.org/10.1042/ebc20180073

Prokaryotes can defend themselves against invading mobile genetic elements (MGEs) by acquiring immune memory against them. The memory is a DNA database located at specific chromosomal sites called CRISPRs (clustered regularly interspaced short palind... Read More about Adaptation processes that build CRISPR immunity: creative destruction, updated.

CRISPR-Cas immunity, DNA repair and genome stability (2018)
Journal Article
Cubbon, A., Ivancic-Bace, I., & Bolt, E. (2018). CRISPR-Cas immunity, DNA repair and genome stability. Bioscience Reports, 38(5), 1-10. https://doi.org/10.1042/bsr20180457

© 2018 The Author(s). Co-opting of CRISPR-Cas ‘Interference’ reactions for editing the genomes of eukaryotic and prokaryotic cells has highlighted crucial support roles for DNA repair systems that strive to maintain genome stability. As front-runners... Read More about CRISPR-Cas immunity, DNA repair and genome stability.

CRISPR-Cas adaptation in Escherichia coli requires RecBCD helicase but not nuclease activity, is independent of homologous recombination, and is antagonized by 5' ssDNA exonucleases (2018)
Journal Article
Radovčić, M., Killelea, T., Savitskaya, E., Wettstein, L., Bolt, E. L., & Ivančić-Baće, I. (2018). CRISPR-Cas adaptation in Escherichia coli requires RecBCD helicase but not nuclease activity, is independent of homologous recombination, and is antagonized by 5' ssDNA exonucleases. Nucleic Acids Research, 46(19), 10173-10183. https://doi.org/10.1093/nar/gky799

© The Author(s) 2018. Prokaryotic adaptive immunity is established against mobile genetic elements (MGEs) by 'naïve adaptation' when DNA fragments from a newly encountered MGE are integrated into CRISPR-Cas systems. In Escherichia coli, DNA integrati... Read More about CRISPR-Cas adaptation in Escherichia coli requires RecBCD helicase but not nuclease activity, is independent of homologous recombination, and is antagonized by 5' ssDNA exonucleases.