Professor Jane A. Grasby
Our laboratory is interested in the interactions and reactions of nucleic acids with a particular interest in enzymes that catalyse phosphoryl transfer reactions in nucleic acids. Most recently the target of our studies has been structure-specific nucleases that recognise aberrant structures in DNA and hydrolyse them to reinstate the genome.
One of our favourite enzymes is flap endonucleases (FENs). FENs are a vital component of the lagging strand DNA replication apparatus in all organisms and also play role in DNA repair in eukaryotes. FENs remove 5’-single-stranded protrusions to double-stranded DNA known as flaps, formed in eukaryotes as a result of DNA polymerase strand displacement synthesis. In humans FENs have to carry out approximately 50 million phosphate diester hydrolyses to allow replication of a single cell. FENs are the prototypical members of a superfamily of structure-specific 5’-nucleases whose differing activities span all major DNA metabolic pathways.
We are working out the molecular mechanism of FENs and related family members, understanding how they recognise DNA substrates and specifically hydrolyse them often using nucleic acid chemistry to help us. With Smythe we are learning how FEN activity is controlled during the cell cycle. Finally, as FENs are overexpressed in certain types of cancer, we are also investigating how to inhibit these enzymes. We also collaborate with Williams on understanding how alkylation of DNA is dealt with in the cell.
- Exell JC, Thompson MJ, Finger LD, Shaw SJ, Debreczeni J, Ward TA, McWhirter C, Siöberg CL, Martinez Molina D, Abbott WM, Jones CD, Nissink JW, Durant ST, Grasby JA (2016) Cellularly Active N-Hydroxyurea FEN1 Inhibitors Block Substrate Entry to the Active Site. Nature Chemical Biology. 12, 815-821.
- Patel N, Exell JC, Jardine E, Ombler B, Finger LD, Ciani B & Grasby JA (2013) Proline Scanning Mutagenesis Reveals a Role for the Flap Endonuclease-1 Helical Cap in Substrate Unpairing. Biol. Chem.. View this article in White Rose Research Online
- Finger LD, Patel N, Beddows A, Ma L, Exell JC, Jardine E, Jones AC & Grasby JA (2013) Observation of unpaired substrate DNA in the flap endonuclease-1 active site.. Nucleic Acids Res, 41(21), 9839-9847. View this article in White Rose Research Online
- Beddows A, Patel N, Finger LD, Atack JM, Williams DM & Grasby JA (2012) Interstrand disulfide crosslinking of DNA bases supports a double nucleotide unpairing mechanism for flap endonucleases.. Chem Commun (Camb), 48(71), 8895-8897.
- Grasby JA, Finger LD, Tsutakawa SE, Atack JM & Tainer JA (2012) Unpairing and gating: Sequence-independent substrate recognition by FEN superfamily nucleases. Trends in Biochemical Sciences, 37(2), 74-84.
- Patel N, Atack JM, Finger LD, Exell JC, Thompson P, Tsutakawa S, Tainer JA, Williams DM & Grasby JA (2012) Flap endonucleases pass 5′-flaps through a flexible arch using a disorder-thread-order mechanism to confer specificity for free 5′-ends.. Nucleic Acids Res, 40(10), 4507-4519.
- Grasby JA, Finger LD, Tsutakawa SE, Atack JM & Tainer JA (2012) Unpairing and gating: sequence-independent substrate recognition by FEN superfamily nucleases.. Trends Biochem Sci, 37(2), 74-84.
- Tsutakawa SE, Classen S, Chapados BR, Arvai AS, Finger LD, Guenther G, Tomlinson CG, Thompson P, Sarker AH, Shen B, Cooper PK, Grasby JA & Tainer JA (2011) Human flap endonuclease structures, DNA double-base flipping, and a unified understanding of the FEN1 superfamily.. Cell, 145(2), 198-211.