Structural basis for FEN-1 substrate specificity and PCNA-mediated activation in DNA replication and repair
- PMID: 14718165
- DOI: 10.1016/s0092-8674(03)01036-5
Structural basis for FEN-1 substrate specificity and PCNA-mediated activation in DNA replication and repair
Abstract
Flap EndoNuclease-1 (FEN-1) and the processivity factor proliferating cell nuclear antigen (PCNA) are central to DNA replication and repair. To clarify the molecular basis of FEN-1 specificity and PCNA activation, we report here structures of FEN-1:DNA and PCNA:FEN-1-peptide complexes, along with fluorescence resonance energy transfer (FRET) and mutational results. FEN-1 binds the unpaired 3' DNA end (3' flap), opens and kinks the DNA, and promotes conformational closing of a flexible helical clamp to facilitate 5' cleavage specificity. Ordering of unstructured C-terminal regions in FEN-1 and PCNA creates an intermolecular beta sheet interface that directly links adjacent PCNA and DNA binding regions of FEN-1 and suggests how PCNA stimulates FEN-1 activity. The DNA and protein conformational changes, composite complex structures, FRET, and mutational results support enzyme-PCNA alignments and a kinked DNA pivot point that appear suitable to coordinate rotary handoffs of kinked DNA intermediates among enzymes localized by the three PCNA binding sites.
Similar articles
-
The interaction site of Flap Endonuclease-1 with WRN helicase suggests a coordination of WRN and PCNA.Nucleic Acids Res. 2005 Dec 2;33(21):6769-81. doi: 10.1093/nar/gki1002. Print 2005. Nucleic Acids Res. 2005. PMID: 16326861 Free PMC article.
-
Structure of the DNA repair and replication endonuclease and exonuclease FEN-1: coupling DNA and PCNA binding to FEN-1 activity.Cell. 1998 Oct 2;95(1):135-46. doi: 10.1016/s0092-8674(00)81789-4. Cell. 1998. PMID: 9778254
-
Physical and functional interaction between human oxidized base-specific DNA glycosylase NEIL1 and flap endonuclease 1.J Biol Chem. 2008 Oct 3;283(40):27028-37. doi: 10.1074/jbc.M802712200. Epub 2008 Jul 28. J Biol Chem. 2008. PMID: 18662981 Free PMC article.
-
The wonders of flap endonucleases: structure, function, mechanism and regulation.Subcell Biochem. 2012;62:301-26. doi: 10.1007/978-94-007-4572-8_16. Subcell Biochem. 2012. PMID: 22918592 Free PMC article. Review.
-
The role of the DNA sliding clamp in Okazaki fragment maturation in archaea and eukaryotes.Biochem Soc Trans. 2011 Jan;39(1):70-6. doi: 10.1042/BST0390070. Biochem Soc Trans. 2011. PMID: 21265749 Review.
Cited by
-
A physiological significance of the functional interaction between Mus81 and Rad27 in homologous recombination repair.Nucleic Acids Res. 2015 Feb 18;43(3):1684-99. doi: 10.1093/nar/gkv025. Epub 2015 Jan 27. Nucleic Acids Res. 2015. PMID: 25628354 Free PMC article.
-
Structural and dynamic determinants of protein-peptide recognition.Structure. 2011 Dec 7;19(12):1837-45. doi: 10.1016/j.str.2011.09.014. Structure. 2011. PMID: 22153506 Free PMC article.
-
Mechanism of DNA substrate recognition by the mammalian DNA repair enzyme, Polynucleotide Kinase.Nucleic Acids Res. 2009 Oct;37(18):6161-73. doi: 10.1093/nar/gkp597. Epub 2009 Aug 11. Nucleic Acids Res. 2009. PMID: 19671525 Free PMC article.
-
Crystallization and preliminary X-ray analysis of flap endonuclease 1 (FEN1) from Desulfurococcus amylolyticus.Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Sep 1;65(Pt 9):923-5. doi: 10.1107/S1744309109031248. Epub 2009 Aug 22. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009. PMID: 19724134 Free PMC article.
-
Crystallization and preliminary crystallographic analysis of the catalytic domain of human flap endonuclease 1 in complex with a nicked DNA product: use of a DPCS kit for efficient protein-DNA complex crystallization.Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Jan 1;64(Pt 1):39-43. doi: 10.1107/S1744309107065372. Epub 2007 Dec 20. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008. PMID: 18097100 Free PMC article.
Publication types
MeSH terms
Substances
Associated data
- Actions
- Actions
- Actions
- Actions
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous
