My laboratory is primarly interested in structure function relationships in proteins which participate in a range of biological processes including DNA modifications, in channeling (in collaboration with Dr S J White, Sheffield) and gamete recognition (in collaboration with Dr C L R Barrett/Prof H D Moore, Sheffield). In addition we are exploiting SELEX (in collaboration with Dr J Grasby, Sheffield) in order to evolve nucleic acids capable of
specific recognition of macromolecular structures.
We have made extensive use of gene fusion technology in order to produce and purify active, recombinant molecules for in vitro analysis. These include the cytosine specific DNA methyltransferases (1,2), the human potassium channel ROMK1 (3) and the human zona pellucida protein ZP3 (4).
Our current work on DNA methyltransferases is concentrating on the analysis of mutants in vivo which exhibit lethal phenotypes as a consequence of their ability to interfere with normal genetic transactions. In addition we are developing vectors encoding the green fluorescent protein for the analysis of ion channel trafficking and gamete recognition.
Using SELEX technology we are currently evolving nucleic acid polymers
which are capable of high affinity recognition of unusual DNA structures to aid our understanding of the origins of the mechanism of nucleic acid recognition.
Figure legend - The modication of DNA by sequence-specific methyltransferases is used strategically by virtually all organisms in the regulation of genetic transactions.
Dickman, M.J., Conroy, M.J., Grasby, J.A. & Hornby, D.P. (2002) RNA Footprinting Analysis using ion pair reverse phase Liquid Chromatography. RNA, 8 247-251
Matin, M.M., Andrews PW. & Hornby, D.P. (2002) Multi-dimensional differential display via ion-pair reversed-phase denaturing
high performance liquid chromatography. Anal. Biochem., 304 47-54
Matin, M.M., Baumer, A. & Hornby, D.P. (2002) An Analytical Method for the Detection of Methylation Differences at Specific
Chromosomal Loci Using Primer Extension and Ion Pair Reverse Phase HPLC. Human Mutation, 20 305-311
Dickman, M.J., Ingleston, S.M., Sedelnikova, S.E., Rafferty, J. B., Lloyd, R.G., Grasby, J.A. & Hornby, D.P. (2002) The RuvABC Resolvasome: Quantitative Analysis of RuvA and RuvC Assembly on Junction DNA. Eur. J. Biochem. 269 5492-5501
Zhou, L., Cheng, X., Connolly, B.A., Dickman, M.J., Hurd, P.J. & Hornby, D.P. (2002) Zebularine: a novel DNA methylation inhibitor that forms a covalent complex with DNA Methyltransferases. J. Mol. Biol. 321 591-599
Ingleston, S.M., Dickman, M.J., Grasby, J.A., Hornby, D.P., Sharples, G.J., & Lloyd R.G. (2002) Holliday junction binding and processing by the RuvA protein of Mycoplasma pneumoniae. Eur J Biochem. 269 1525-33.
Harris, V.H., Smith C.L. Cummins, W.J., Hamilton, A.L., Adams, H., Dickman M., Hornby, D.P. & Williams D.M. (2003) The effect of Tautomeric Constant on the Specificity of Nucleotide Incorporation
during DNA Replication: Support for the Rare Tautomer Hypothesis of Substitution Mutagenesis. J.Mol.Biol. 326, 1389-1401.
