2qm4

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==Overview==
==Overview==
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The recently characterised 299-residue human XLF/Cernunnos protein plays a, crucial role in DNA repair by non-homologous end joining (NHEJ) and, interacts with the XRCC4-DNA Ligase IV complex. Here, we report the, crystal structure of the XLF (1-233) homodimer at 2.3 A resolution, confirming the predicted structural similarity to XRCC4. The XLF, coiled-coil, however, is shorter than that of XRCC4 and undergoes an, unexpected reverse in direction giving rise to a short distorted four, helical bundle and a C-terminal helical structure wedged between the, coiled-coil and head domain. The existence of a dimer as the major species, is confirmed by size-exclusion chromatography, analytical, ultracentrifugation, small-angle X-ray scattering and other biophysical, methods. We show that the XLF structure is not easily compatible with a, proposed XRCC4:XLF heterodimer. However, we demonstrate interactions, between dimers of XLF and XRCC4 by surface plasmon resonance and analyse, these in terms of surface properties, amino-acid conservation and, mutations in immunodeficient patients. Our data are most consistent with, head-to-head interactions in a 2:2:1 XRCC4:XLF:Ligase IV complex.
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The recently characterised 299-residue human XLF/Cernunnos protein plays a crucial role in DNA repair by non-homologous end joining (NHEJ) and interacts with the XRCC4-DNA Ligase IV complex. Here, we report the crystal structure of the XLF (1-233) homodimer at 2.3 A resolution, confirming the predicted structural similarity to XRCC4. The XLF coiled-coil, however, is shorter than that of XRCC4 and undergoes an unexpected reverse in direction giving rise to a short distorted four helical bundle and a C-terminal helical structure wedged between the coiled-coil and head domain. The existence of a dimer as the major species is confirmed by size-exclusion chromatography, analytical ultracentrifugation, small-angle X-ray scattering and other biophysical methods. We show that the XLF structure is not easily compatible with a proposed XRCC4:XLF heterodimer. However, we demonstrate interactions between dimers of XLF and XRCC4 by surface plasmon resonance and analyse these in terms of surface properties, amino-acid conservation and mutations in immunodeficient patients. Our data are most consistent with head-to-head interactions in a 2:2:1 XRCC4:XLF:Ligase IV complex.
==About this Structure==
==About this Structure==
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==Reference==
==Reference==
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Crystal structure of human XLF/Cernunnos reveals unexpected differences from XRCC4 with implications for NHEJ., Li Y, Chirgadze DY, Bolanos-Garcia VM, Sibanda BL, Davies OR, Ahnesorg P, Jackson SP, Blundell TL, EMBO J. 2007 Nov 29;. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=18046455 18046455]
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Crystal structure of human XLF/Cernunnos reveals unexpected differences from XRCC4 with implications for NHEJ., Li Y, Chirgadze DY, Bolanos-Garcia VM, Sibanda BL, Davies OR, Ahnesorg P, Jackson SP, Blundell TL, EMBO J. 2008 Jan 9;27(1):290-300. Epub 2007 Nov 29. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=18046455 18046455]
[[Category: Homo sapiens]]
[[Category: Homo sapiens]]
[[Category: Single protein]]
[[Category: Single protein]]
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[[Category: Blundell, T.L.]]
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[[Category: Blundell, T L.]]
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[[Category: Bolanos-Garcia, V.M.]]
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[[Category: Bolanos-Garcia, V M.]]
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[[Category: Chirgadze, D.Y.]]
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[[Category: Chirgadze, D Y.]]
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[[Category: Davies, O.R.]]
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[[Category: Davies, O R.]]
[[Category: Li, Y.]]
[[Category: Li, Y.]]
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[[Category: Sibanda, B.L.]]
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[[Category: Sibanda, B L.]]
[[Category: beta-sandwich]]
[[Category: beta-sandwich]]
[[Category: coiled-coil]]
[[Category: coiled-coil]]
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[[Category: xrcc4 like factor]]
[[Category: xrcc4 like factor]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Jan 31 10:58:57 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:40:24 2008''

Revision as of 16:40, 21 February 2008


2qm4, resolution 2.30Å

Drag the structure with the mouse to rotate

Crystal structure of human XLF/Cernunnos, a non-homologous end-joining factor

Overview

The recently characterised 299-residue human XLF/Cernunnos protein plays a crucial role in DNA repair by non-homologous end joining (NHEJ) and interacts with the XRCC4-DNA Ligase IV complex. Here, we report the crystal structure of the XLF (1-233) homodimer at 2.3 A resolution, confirming the predicted structural similarity to XRCC4. The XLF coiled-coil, however, is shorter than that of XRCC4 and undergoes an unexpected reverse in direction giving rise to a short distorted four helical bundle and a C-terminal helical structure wedged between the coiled-coil and head domain. The existence of a dimer as the major species is confirmed by size-exclusion chromatography, analytical ultracentrifugation, small-angle X-ray scattering and other biophysical methods. We show that the XLF structure is not easily compatible with a proposed XRCC4:XLF heterodimer. However, we demonstrate interactions between dimers of XLF and XRCC4 by surface plasmon resonance and analyse these in terms of surface properties, amino-acid conservation and mutations in immunodeficient patients. Our data are most consistent with head-to-head interactions in a 2:2:1 XRCC4:XLF:Ligase IV complex.

About this Structure

2QM4 is a Single protein structure of sequence from Homo sapiens. Full crystallographic information is available from OCA.

Reference

Crystal structure of human XLF/Cernunnos reveals unexpected differences from XRCC4 with implications for NHEJ., Li Y, Chirgadze DY, Bolanos-Garcia VM, Sibanda BL, Davies OR, Ahnesorg P, Jackson SP, Blundell TL, EMBO J. 2008 Jan 9;27(1):290-300. Epub 2007 Nov 29. PMID:18046455

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