Files in this item
PCNA stimulates catalysis by structure-specific nucleases using two distinct mechanisms : substrate targeting and catalytic step
Item metadata
dc.contributor.author | Hutton, R D | |
dc.contributor.author | Roberts, J A | |
dc.contributor.author | Penedo, Carlos | |
dc.contributor.author | White, Malcolm F | |
dc.date.accessioned | 2011-12-12T12:35:44Z | |
dc.date.available | 2011-12-12T12:35:44Z | |
dc.date.issued | 2008-12 | |
dc.identifier | 424475 | |
dc.identifier | effd0707-488a-4d59-89d6-8965cfae8253 | |
dc.identifier | 000261299700006 | |
dc.identifier | 57149118629 | |
dc.identifier.citation | Hutton , R D , Roberts , J A , Penedo , C & White , M F 2008 , ' PCNA stimulates catalysis by structure-specific nucleases using two distinct mechanisms : substrate targeting and catalytic step ' , Nucleic Acids Research , vol. 36 , no. 21 , pp. 6720-6727 . https://doi.org/10.1093/nar/gkn745 | en |
dc.identifier.issn | 0305-1048 | |
dc.identifier.other | ORCID: /0000-0003-1543-9342/work/47136082 | |
dc.identifier.other | ORCID: /0000-0002-5807-5385/work/74872771 | |
dc.identifier.uri | https://hdl.handle.net/10023/2103 | |
dc.description | This work was supported by the Biotechnology and Biological Sciences Research Council [grants BBD0014391 and BBE0146741] | en |
dc.description.abstract | The sliding clamp Proliferating Cell Nuclear Antigen (PCNA) functions as a recruiter and organizer of a wide variety of DNA modifying enzymes including nucleases, helicases, polymerases and glycosylases. The 5-flap endonuclease Fen-1 is essential for Okazaki fragment processing in eukaryotes and archaea, and is targeted to the replication fork by PCNA. Crenarchaeal XPF, a 3-flap endonuclease, is also stimulated by PCNA in vitro. Using a novel continuous fluorimetric assay, we demonstrate that PCNA activates these two nucleases by fundamentally different mechanisms. PCNA stimulates Fen-1 by increasing the enzymes binding affinity for substrates, as suggested previously. However, PCNA activates XPF by increasing the catalytic rate constant by four orders of magnitude without affecting the K-M. PCNA may function as a platform upon which XPF exerts force to distort DNA substrates, destabilizing the substrate and/or stabilizing the transition state structure. This suggests that PCNA can function directly in supporting catalysis as an essential cofactor in some circumstances, a new role for a protein that is generally assumed to perform a passive targeting and organizing function in molecular biology. This could provide a mechanism for the exquisite control of nuclease activity targeted to specific circumstances, such as replication forks or damaged DNA with pre-loaded PCNA. | |
dc.format.extent | 8 | |
dc.format.extent | 4394801 | |
dc.language.iso | eng | |
dc.relation.ispartof | Nucleic Acids Research | en |
dc.subject | Heterotrimeric PCNA | en |
dc.subject | Mismatch repair | en |
dc.subject | Sulfolobus-solfataricus | en |
dc.subject | DNA | en |
dc.subject | Endonuclease | en |
dc.subject | Proteins | en |
dc.subject | Ubiquitin | en |
dc.subject | Enzyme | en |
dc.subject | Sumo | en |
dc.title | PCNA stimulates catalysis by structure-specific nucleases using two distinct mechanisms : substrate targeting and catalytic step | en |
dc.type | Journal article | en |
dc.contributor.sponsor | BBSRC | en |
dc.contributor.sponsor | BBSRC | en |
dc.contributor.sponsor | BBSRC | en |
dc.contributor.institution | University of St Andrews. School of Biology | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.identifier.doi | 10.1093/nar/gkn745 | |
dc.description.status | Peer reviewed | en |
dc.identifier.url | http://www.scopus.com/inward/record.url?scp=57149118629&partnerID=8YFLogxK | en |
dc.identifier.grantnumber | BB/E014674/1 | en |
dc.identifier.grantnumber | BB/E014674/1 | en |
dc.identifier.grantnumber | BB/D001439/1 | en |
This item appears in the following Collection(s)
Items in the St Andrews Research Repository are protected by copyright, with all rights reserved, unless otherwise indicated.