Ed Bolt

Contact

• workRoom D42 The University of Nottingham Medical School
  Queen's Medical Centre
  Nottingham
  NG7 2UH
  UK
• work0115 82 30194
• fax0115 82 30142
• ed.bolt@nottingham.ac.uk

Teaching Summary

Structure and function of protein-nucleic acid interactions

Genome Dynamics

Molecular Microbiology

Research Summary

Genome dynamics and DNA repair

Our group researches two major themes central to the genomes of bacteria and archaea; (i) genome plasticity, and (ii) repair and recovery of DNA replication.

(i). Our research on genome plasticity highlights the functions of cas genes, which are highly conserved in about 50% of known prokaryotic species and are associated with CRISPR repeat DNA elements. Cas-CRISPR seems to have multifarious roles feeding into several aspects of cellular metabolism. We are currently investigating cas-CRISPR using bacterial genetics and in vitro biochemistry.

(ii). we are also interested in how DNA helicases influence repair of broken DNA replication forks, especially through homologous recombination. This project focuses on the actions of Hel308 DNA helicase, which seems to interact with replication forks. We are using biochemistry of archaeal Hel308 and its human homologue to work out the helicase mechanism and interactions with other proteins.

Selected Publications

• HOWARD, J.L, DELMAS, S, IVANCIC-BACE, I and BOLT, E.L, 2011. Helicase dissociation and annealing of RNA-DNA hybrids by Escherichia coli Cas3 protein Biochemical Journal. doi:10.1042/BJ20110901,
• WOODMAN, ISABEL L, BRAMMER, KIRSTY and BOLT, EDWARD L, 2011. Physical interaction between archaeal DNA repair helicase Hel308 and Replication Protein A (RPA). DNA repair. 10(3), 306-13
• WOODMAN, ISABEL L and BOLT, EDWARD L, 2011. Winged helix domains with unknown function in Hel308 and related helicases. Biochemical Society transactions. 39(1), 140-4
• MATILLA, INMACULADA, ALFONSO, CARLOS, RIVAS, GERMAN, BOLT, EDWARD L, DE LA CRUZ, FERNANDO and CABEZON, ELENA, 2010. The conjugative DNA translocase TrwB is a structure-specific DNA-binding protein. The Journal of biological chemistry. 285(23), 17537-44

• View all publications

• HOWARD, J.L, DELMAS, S, IVANCIC-BACE, I and BOLT, E.L, 2011. Helicase dissociation and annealing of RNA-DNA hybrids by Escherichia coli Cas3 protein Biochemical Journal. doi:10.1042/BJ20110901,
• WOODMAN, ISABEL L, BRAMMER, KIRSTY and BOLT, EDWARD L, 2011. Physical interaction between archaeal DNA repair helicase Hel308 and Replication Protein A (RPA). DNA repair. 10(3), 306-13
• WOODMAN, ISABEL L and BOLT, EDWARD L, 2011. Winged helix domains with unknown function in Hel308 and related helicases. Biochemical Society transactions. 39(1), 140-4
• MATILLA, INMACULADA, ALFONSO, CARLOS, RIVAS, GERMAN, BOLT, EDWARD L, DE LA CRUZ, FERNANDO and CABEZON, ELENA, 2010. The conjugative DNA translocase TrwB is a structure-specific DNA-binding protein. The Journal of biological chemistry. 285(23), 17537-44
• MACHWE,, XIAO,, LLOYD,, BOLT, and ORREN,, 2007. Replication fork regression in vitro by the Werner syndrome protein (WRN): Holliday junction formation, the effect of leading arm structure and a potential role for WRN exonuclease activity. Nucleic Acids Research. 35(17), 5729-47
• WOODMAN, ISABEL L, BRIGGS, GEOFFREY S and BOLT, EDWARD L, 2007. Archaeal Hel308 domain V couples DNA binding to ATP hydrolysis and positions DNA for unwinding over the helicase ratchet. Journal Molecular Biology. 374(5), 1139-44
• MACMASTER, R, SEDELNIKOVA, S, BAKER, P.J, BOLT, E.L, LLOYD, R.G and RAFFERTY, J.B., 2006. RusA Holliday junction resolvase: DNA complex structure—insights into selectivity and specificity Nucleic Acids Research. 34(19), 5577–5584
• GUY, C.P., HALDENBY, S., BRINDLEY, A., WALSH, D.A., BRIGGS, G.S., WARREN, M.J., ALLERS, T. and BOLT, E.L., 2006. Interactions of RadB, a DNA repair protein in archaea, with DNA and ATP Journal of Molecular Biology. 358(1), 46-56
• GUY, C.P. and BOLT, E.L., 2005. Archaeal Hel308 helicase targets replication forks <i>in vivo</i> and <i>in vitro</i> and unwinds lagging strands Nucleic Acids Research. 33(11), 3678-3690
• BOLT, E. L., 2005. Helicases that interact with replication forks: new candidates from archaea Biochemical Society Transactions. VOL 33(PART 6), 1471-1473
• GUY, C.P., MAJERNIK, A.I., CHONG, J.P.J. and BOLT, E.L., 2004. A novel nuclease-ATPase (Nar71) from archaea is part of a proposed thermophilic DNA repair system Nucleic Acids Research. 32(21), 6176-6186
• SHARPLES, G. J., CURTIS, F. A., MCGLYNN, P. and BOLT, E. L., 2004. Holliday Junction Binding and Resolution by the Rap Structure-specific Endonuclease of Phage l Journal of Molecular Biology. VOL 340(NUMBER 4), 739-751
• RAFFERTY, J.B., BOLT, E.L., MURANOVA, T.A., SEDELNIKOVA, S.E., LEONARD, P., PASQUO, A., BAKER, P.J., RICE, D.W., SHARPLES, G.J. and LLOYD, R.G., 2003. The structure of Escherichia coli RusA endonuclease reveals a new Holliday junction DNA binding fold Structure. 11(12), 1557-1567
• MURANOVA, T A., SEDELNIKOVA, S.E., LEONARD, P.M., PASQUO, A., BOLT, E.L., LLOYD, R.G. and RAFFERTY, J.B., 2003. Crystallization of RusA Holliday junction resolvase from Escherichia coli Acta Crystallographica. Section D: Biological Crystallography. 59(12), 2262-2264
• BOLT, E. L. and GUY, C. P., 2003. Homologous recombination in Archaea: new Holliday junction helicases Biochemical Society Transactions. VOL 31(PART 3), 703-705
• BOLT, E.L. and LLOYD, R.G., 2002. Substrate specificity of RusA resolvase reveals the DNA structures targeted by RuvAB and RecG in vivo Molecular Cell. 10(1), 187-198
• SHARPLES, G.J., BOLT, E.L. and LLOYD, R.G., 2002. RusA proteins from the extreme thermophile <i>Aquifex aeolicus</i> and lactococcal phage r1t resolve Holliday junctions Molecular Microbiology. 44(2), 549-560
• BOLT, E.L., LLOYD, R.G. and SHARPLES, G.J., 2001. Genetic analysis of an archaeal Holliday junction resolvase in <i>Escherichia coli</i> Journal of Molecular Biology. 310(3), 577-589