Papers by J. N Mark Glover
Structural insights into DNA double-strand break signaling
Biochemical Journal
Genomic integrity is most threatened by double-strand breaks, which, if left unrepaired, lead to ... more Genomic integrity is most threatened by double-strand breaks, which, if left unrepaired, lead to carcinogenesis or cell death. The cell generates a network of protein–protein signaling interactions that emanate from the DNA damage which are now recognized as a rich basis for anti-cancer therapy development. Deciphering the structures of signaling proteins has been an uphill task owing to their large size and complex domain organization. Recent advances in mammalian protein expression/purification and cryo-EM-based structure determination have led to significant progress in our understanding of these large multidomain proteins. This review is an overview of the structural principles that underlie some of the key signaling proteins that function at the double-strand break site. We also discuss some plausible ideas that could be considered for future structural approaches to visualize and build a more complete understanding of protein dynamics at the break site.
CSMCB/SCBBMC BULLETIN 2005 45 Structural Basis for Phosphorylation- dependent Signaling in the DNA Damage Response
The response of eukaryotic cells to DNA damage requires a multitude of protein-protein interactio... more The response of eukaryotic cells to DNA damage requires a multitude of protein-protein interactions that mediate the ordered repair of the damage and arrest of the cell cycle until repair is complete. Two conserved protein modules, BRCT and FHA domains, play key roles in the DNA damage response as recognition elements for nuclear Ser/Thr phosphorylation induced by DNA dam-age-responsive kinases. BRCT domains, first iden-tified at the C-terminus of BRCA1, often occur as multiple tandem repeats of individual BRCT mod-ules. Our recent structural and functional work has revealed how BRCT repeats recognize phospho-serine protein targets, and have revealed a second-ary binding pocket at the interface between tan-dem repeats that recognizes the amino acid 3 residues C-terminal to the phospho-serine. We have also studied the molecular function of the FHA domain of the DNA repair enzyme, PNK. This domain interacts with threonine-phosphory-lated XRCC1 and XRCC4, which is responsible for the r...
Revealed in the Structural of Ndt80-MSE DNA Comple
by
Neurological disorders associated with DNA strand-break processing enzymes
Mechanisms of Ageing and Development, 2017
The termini of DNA strand breaks induced by reactive oxygen species or by abortive DNA metabolic ... more The termini of DNA strand breaks induced by reactive oxygen species or by abortive DNA metabolic intermediates require processing to enable subsequent gap filling and ligation to proceed. The three proteins, tyrosyl DNA-phosphodiesterase 1 (TDP1), aprataxin (APTX) and polynucleotide kinase/phosphatase (PNKP) each act on a discrete set of modified strand-break termini. Recently, a series of neurodegenerative and neurodevelopmental disorders have been associated with mutations in the genes coding for these proteins. Mutations in TDP1 and APTX have been linked to Spinocerebellar ataxia with axonal neuropathy (SCAN1) and Ataxia-ocular motor apraxia 1 (AOA1), respectively, while mutations in PNKP are considered to be responsible for Microcephaly with seizures (MCSZ) and Ataxia-ocular motor apraxia 4 (AOA4). Here we present an overview of the mechanisms of these proteins and how their impairment may give rise to their respective disorders.
F1000 - Post-publication peer review of the biomedical literature, 2012
Poly(ADP-ribose) polymerase I (PARP1) is a primary DNA damage sensor whose (ADP-ribose) polymeras... more Poly(ADP-ribose) polymerase I (PARP1) is a primary DNA damage sensor whose (ADP-ribose) polymerase activity is acutely regulated by interaction with DNA breaks. Upon activation at sites of DNA damage, PARP1 modifies itself and other proteins by covalent addition of long branched polymers of ADP-ribose, which in turn recruit downstream DNA repair and chromatin remodelling factors. PARP1 recognizes DNA damage through its N-terminal DNA-binding domain (DBD), which consists of a tandem repeat of an unusual zinc-finger (ZnF) domain. We have now determined the crystal structure of the human PARP1-DBD bound to a DNA break. Along with functional analysis of PARP1 recruitment to sites of DNA damage in vivo, the structure reveals a dimeric assembly whereby ZnF1 and ZnF2 domains from separate PARP1 molecules form a strand-break recognition module that helps activate PARP1 by facilitating its dimerization and consequent trans-automodification. Short-patch repair of DNA single-strand breaks is initiated by poly(ADP-ribose) polymerase-1 (PARP1) -a multi-domain enzyme activated by binding of its N-terminal DNA-binding domain (DBD) to DNA breaks 1-5 .
F1000 - Post-publication peer review of the biomedical literature, 2010
Author contribution: J.S. helped characterize MCSZ syndrome, identified MCSZ locus and calculated... more Author contribution: J.S. helped characterize MCSZ syndrome, identified MCSZ locus and calculated LOD scores, sequenced genes in MCSZ locus to identify PNKP mutations, wrote the manuscript, E.C.G. helped characterize MCSZ syndrome, identified moderately affected MCSZ family, performed RT-PCR on moderately affected family, performed comet assays, organized and analyzed sequenom experiments, did analysis of PNKP mutation, performed mouse RNAi experiments, helped perform mouse in situs, wrote the manuscript, C.A.M. sequenced genes in MCSZ locus to identify PNKP mutations and helped perform human in situs, M.H. identified patients and provided clinical information, J.J.R. performed PNKP Western blot and confirmatory comet assays, W.E. identified patients and provided clinical information, A.B. organized clinical information and patient samples, B.B. organized clinical information and patient samples, D.G. organized patient samples and helped perform sequenome experiments, K.A. organized patient samples and helped perform sequencing experiments, V.S.G. helped analyze sequenom experiments, B.S.C. helped organize clinical information to identify MCSZ syndrome, A.G. identified patients and provided clinical information, R.S.H. helped organize genetic data and calculate LOD scores, M.T. identified patients and provided clinical information, K.W.C. advised on comet assays, supervised PNKP Western blot and edited manuscript, A.J.B. characterized MRIs for patient classification, C.A.W. directed overall research and wrote the manuscript.
Faculty of 1000 evaluation for Discovery of Cell-Permeable Inhibitors That Target the BRCT Domain of BRCA1 Protein by Using Small-Molecule Microarray
F1000 - Post-publication peer review of the biomedical literature, 2014
Nucleic Acids Research, 2011
The conjugative transfer of F-like plasmids such as F, R1, R100 and pED208, between bacterial cel... more The conjugative transfer of F-like plasmids such as F, R1, R100 and pED208, between bacterial cells requires TraM, a plasmid-encoded DNA-binding protein. TraM tetramers bridge the origin of transfer (oriT) to a key component of the conjugative pore, the coupling protein TraD. Here we show that TraM recognizes a high-affinity DNA-binding site, sbmA, as a cooperative dimer of tetramers. The crystal structure of the TraM-sbmA complex from the plasmid pED208 shows that binding cooperativity is mediated by DNA kinking and unwinding, without any direct contact between tetramers. Sequence-specific DNA recognition is carried out by TraM's N-terminal ribbon-helix-helix (RHH) domains, which bind DNA in a staggered arrangement. We demonstrate that both DNA-binding specificity, as well as selective interactions between TraM and the C-terminal tail of its cognate TraD mediate conjugation specificity within the F-like family of plasmids. The ability of TraM to cooperatively bind DNA without interaction between tetramers leaves the C-terminal TraM tetramerization domains free to make multiple interactions with TraD, driving recruitment of the plasmid to the conjugative pore.
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Papers by J. N Mark Glover