title: Crystallization and preliminary X-ray diffraction analysis of the human XRCC4-XLF complex.
abstract: XRCC4 and XLF are key proteins in the repair of DNA double-strand breaks through nonhomologous end-joining. Together, they form a complex that stimulates the ligation of double-strand breaks. Owing to the suggested filamentous nature of this complex, structural studies via X-ray crystallography have proven difficult. Multiple truncations of the XLF and XRCC4 proteins were cocrystallized, but yielded low-resolution diffraction (~20 Å). However, a combination of microseeding, dehydration and heavy metals improved the diffraction of XRCC4(Δ157)-XLF(Δ224) crystals to 3.9 Å resolution. Although molecular replacement alone was unable to produce a solution, when combined with the anomalous signal from tantalum bromide clusters initial phasing was successfully obtained.
authors.affiliation: Biochemistry and Biomedical Sciences, McMaster University, 1200 Main Street West, Hamilton, Ontario L8N 3Z5, Canada.
authors.forename: Sara N
authors.initials: SN
authors.lastname: Andres
authors.forename: Murray S
authors.initials: MS
authors.lastname: Junop
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D045643:DNA Repair Enzymes / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D011485:Protein Binding
chemical_list: D004268:DNA-Binding Proteins; C508551:NHEJ1 protein, human; C101467:XRCC4 protein, human; D045643:DNA Repair Enzymes
references.citation: Am J Hum Genet. 1990 Sep;47(3):459-69
references.pmid: 1697445
references.citation: EMBO J. 1999 Apr 1;18(7):2008-18
references.pmid: 10202163
references.citation: Proc Natl Acad Sci U S A. 2007 Mar 13;104(11):4518-23
references.pmid: 17360556
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Structure. 2003 Feb;11(2):139-45
references.pmid: 12575933
references.citation: Mol Cell. 2007 Dec 28;28(6):1093-101
references.pmid: 18158905
references.citation: EMBO J. 2008 Jan 9;27(1):290-300
references.pmid: 18046455
references.citation: Proc Natl Acad Sci U S A. 2007 May 8;104(19):7851-6
references.pmid: 17470781
references.citation: J Mol Biol. 1997 Jul 4;270(1):1-7
references.pmid: 9231895
references.citation: Structure. 2010 Nov 10;18(11):1431-42
references.pmid: 21070942
references.citation: Nature. 2009 Mar 26;458(7237):475-80
references.pmid: 19325628
references.citation: J Biol Chem. 2011 Sep 16;286(37):32638-50
references.pmid: 21775435
references.citation: EMBO J. 2000 Nov 15;19(22):5962-70
references.pmid: 11080143
references.citation: J Biol Chem. 2010 Aug 20;285(34):26475-83
references.pmid: 20558749
references.citation: Mol Cell Biol. 2009 Jun;29(11):3163-72
references.pmid: 19332554
references.citation: Subcell Biochem. 2010;50:279-96
references.pmid: 20012587
references.citation: Nature. 1999 Aug 26;400(6747):841-7
references.pmid: 10476961
references.citation: Cell. 2006 Jan 27;124(2):301-13
references.pmid: 16439205