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| | ==Crystal structure of paFAN1 bound to 2nt 5'flap DNA with gap== | | ==Crystal structure of paFAN1 bound to 2nt 5'flap DNA with gap== |
| - | <StructureSection load='5y7q' size='340' side='right' caption='[[5y7q]], [[Resolution|resolution]] 2.70Å' scene=''> | + | <StructureSection load='5y7q' size='340' side='right'caption='[[5y7q]], [[Resolution|resolution]] 2.70Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5y7q]] is a 4 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5Y7Q OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5Y7Q FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5y7q]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_aeruginosa_PAO1 Pseudomonas aeruginosa PAO1] and [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5Y7Q OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5Y7Q FirstGlance]. <br> |
| - | </td></tr><tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Phosphodiesterase_I Phosphodiesterase I], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.4.1 3.1.4.1] </span></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.7Å</td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5y7q FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5y7q OCA], [http://pdbe.org/5y7q PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5y7q RCSB], [http://www.ebi.ac.uk/pdbsum/5y7q PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5y7q ProSAT]</span></td></tr> | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5y7q FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5y7q OCA], [https://pdbe.org/5y7q PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5y7q RCSB], [https://www.ebi.ac.uk/pdbsum/5y7q PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5y7q ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/FAN1_PSEAE FAN1_PSEAE]] Nuclease required for the repair of DNA interstrand cross-links (ICL). Acts as a 5'-3' exonuclease that anchors at a cut end of DNA and cleaves DNA successively at every third nucleotide, allowing to excise an ICL from one strand through flanking incisions. Also has endonuclease activity toward 5'-flaps (PubMed:25319828).[UniProtKB:Q9Y2M0]<ref>PMID:24981866</ref> <ref>PMID:25319828</ref> | + | [https://www.uniprot.org/uniprot/FAN1_PSEAE FAN1_PSEAE] Nuclease required for the repair of DNA interstrand cross-links (ICL). Acts as a 5'-3' exonuclease that anchors at a cut end of DNA and cleaves DNA successively at every third nucleotide, allowing to excise an ICL from one strand through flanking incisions. Also has endonuclease activity toward 5'-flaps (PubMed:25319828).[UniProtKB:Q9Y2M0]<ref>PMID:24981866</ref> <ref>PMID:25319828</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Phosphodiesterase I]] | + | [[Category: Large Structures]] |
| - | [[Category: Cho, Y]] | + | [[Category: Pseudomonas aeruginosa PAO1]] |
| - | [[Category: Jin, H]] | + | [[Category: Synthetic construct]] |
| - | [[Category: Hydrolase-dna complex]] | + | [[Category: Cho Y]] |
| - | [[Category: Nuclease]] | + | [[Category: Jin H]] |
| Structural highlights
Function
FAN1_PSEAE Nuclease required for the repair of DNA interstrand cross-links (ICL). Acts as a 5'-3' exonuclease that anchors at a cut end of DNA and cleaves DNA successively at every third nucleotide, allowing to excise an ICL from one strand through flanking incisions. Also has endonuclease activity toward 5'-flaps (PubMed:25319828).[UniProtKB:Q9Y2M0][1] [2]
Publication Abstract from PubMed
DNA interstrand cross-links (ICLs) block the progress of the replication and transcription machineries and can weaken chromosomal stability, resulting in various diseases. FANCD2-FANCI-associated nuclease (FAN1) is a conserved structure-specific nuclease that unhooks DNA ICLs independently of the Fanconi anemia pathway. Recent structural studies have proposed two different mechanistic features for ICL unhooking by human FAN1: a specific basic pocket that recognizes the terminal phosphate of a 1-nucleotide (nt) 5' flap or FAN1 dimerization. Herein, we show that despite lacking these features, Pseudomonas aeruginosa FAN1 (PaFAN1) cleaves substrates at approximately 3-nt intervals and resolves ICLs. Crystal structures of PaFAN1 bound to various DNA substrates revealed that its conserved basic Arg/Lys patch comprising Arg-228 and Lys-260 recognizes phosphate groups near the 5' terminus of a DNA substrate with a 1-nt flap or a nick. Substitution of Lys-260 did not affect PaFAN1's initial endonuclease activity but significantly decreased its subsequent exonuclease activity and ICL unhooking. The Arg/Lys patch also interacted with phosphates at a 3-nt gap, and this interaction could drive movement of the scissile phosphates into the PaFAN1-active site. In human FAN1, the ICL-resolving activity was not affected by individual disruption of the Arg/Lys patch or basic pocket. However, simultaneous substitution of both FAN1 regions significantly reduced its ICL-resolving activity, suggesting that these two basic regions play a complementary role in ICL repair. On the basis of these findings, we propose a conserved role for two basic regions in FAN1 to guide ICL unhooking and to maintain genomic stability.
Structural mechanism of DNA interstrand cross-link unhooking by the bacterial FAN1 nuclease.,Jin H, Roy U, Lee G, Scharer OD, Cho Y J Biol Chem. 2018 Apr 27;293(17):6482-6496. doi: 10.1074/jbc.RA118.002171. Epub, 2018 Mar 7. PMID:29514982[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Pennell S, Declais AC, Li J, Haire LF, Berg W, Saldanha JW, Taylor IA, Rouse J, Lilley DM, Smerdon SJ. FAN1 activity on asymmetric repair intermediates is mediated by an atypical monomeric virus-type replication-repair nuclease domain. Cell Rep. 2014 Jul 10;8(1):84-93. doi: 10.1016/j.celrep.2014.06.001. Epub 2014, Jun 26. PMID:24981866 doi:http://dx.doi.org/10.1016/j.celrep.2014.06.001
- ↑ Gwon GH, Kim Y, Liu Y, Watson AT, Jo A, Etheridge TJ, Yuan F, Zhang Y, Kim Y, Carr AM, Cho Y. Crystal structure of a Fanconi anemia-associated nuclease homolog bound to 5' flap DNA: basis of interstrand cross-link repair by FAN1. Genes Dev. 2014 Oct 15;28(20):2276-90. doi: 10.1101/gad.248492.114. PMID:25319828 doi:http://dx.doi.org/10.1101/gad.248492.114
- ↑ Jin H, Roy U, Lee G, Scharer OD, Cho Y. Structural mechanism of DNA interstrand cross-link unhooking by the bacterial FAN1 nuclease. J Biol Chem. 2018 Apr 27;293(17):6482-6496. doi: 10.1074/jbc.RA118.002171. Epub, 2018 Mar 7. PMID:29514982 doi:http://dx.doi.org/10.1074/jbc.RA118.002171
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