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Methods Mol Biol
2022 Jan 01;2444:243-269. doi: 10.1007/978-1-0716-2063-2_15.
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Purification and Characterization of Human DNA Ligase IIIα Complexes After Expression in Insect Cells.
Rashid I
,
Tsai MS
,
Sverzhinsky A
,
Hlaing AS
,
Shih B
,
Thwin AC
,
Lin JG
,
Maw SS
,
Pascal JM
,
Tomkinson AE
.
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With improvements in biophysical approaches, there is growing interest in characterizing large, flexible multi-protein complexes. The use of recombinant baculoviruses to express heterologous genes in cultured insect cells has advantages for the expression of human protein complexes because of the ease of co-expressing multiple proteins in insect cells and the presence of a conserved post-translational machinery that introduces many of the same modifications found in human cells. Here we describe the preparation of recombinant baculoviruses expressing DNA ligase IIIα, XRCC1, and TDP1, their subsequent co-expression in cultured insect cells, the purification of complexes containing DNA ligase IIIα from insect cell lysates, and their characterization by multi-angle light scattering linked to size exclusion chromatography and negative stain electron microscopy.
Abdella,
Structure of the human Mediator-bound transcription preinitiation complex.
2021, Pubmed
Abdella,
Structure of the human Mediator-bound transcription preinitiation complex.
2021,
Pubmed
Caldecott,
An interaction between the mammalian DNA repair protein XRCC1 and DNA ligase III.
1994,
Pubmed
,
Xenbase
Caldecott,
XRCC1 protein; Form and function.
2019,
Pubmed
Cannan,
The Human Ligase IIIα-XRCC1 Protein Complex Performs DNA Nick Repair after Transient Unwrapping of Nucleosomal DNA.
2017,
Pubmed
Chen,
Structural basis of long-range to short-range synaptic transition in NHEJ.
2021,
Pubmed
Cheng,
A primer to single-particle cryo-electron microscopy.
2015,
Pubmed
Della-Maria,
Human Mre11/human Rad50/Nbs1 and DNA ligase IIIalpha/XRCC1 protein complexes act together in an alternative nonhomologous end joining pathway.
2011,
Pubmed
,
Xenbase
El-Khamisy,
Defective DNA single-strand break repair in spinocerebellar ataxia with axonal neuropathy-1.
2005,
Pubmed
,
Xenbase
Ellenberger,
Eukaryotic DNA ligases: structural and functional insights.
2008,
Pubmed
Gallagher,
Negative-Stain Transmission Electron Microscopy of Molecular Complexes for Image Analysis by 2D Class Averaging.
2019,
Pubmed
Gao,
DNA ligase III is critical for mtDNA integrity but not Xrcc1-mediated nuclear DNA repair.
2011,
Pubmed
,
Xenbase
Gradia,
MacroBac: New Technologies for Robust and Efficient Large-Scale Production of Recombinant Multiprotein Complexes.
2017,
Pubmed
Hammel,
Ku and DNA-dependent protein kinase dynamic conformations and assembly regulate DNA binding and the initial non-homologous end joining complex.
2010,
Pubmed
Hammel,
An Intrinsically Disordered APLF Links Ku, DNA-PKcs, and XRCC4-DNA Ligase IV in an Extended Flexible Non-homologous End Joining Complex.
2016,
Pubmed
Hammel,
An atypical BRCT-BRCT interaction with the XRCC1 scaffold protein compacts human DNA Ligase IIIα within a flexible DNA repair complex.
2021,
Pubmed
Huang,
TDP1 repairs nuclear and mitochondrial DNA damage induced by chain-terminating anticancer and antiviral nucleoside analogs.
2013,
Pubmed
Lakshmipathy,
The human DNA ligase III gene encodes nuclear and mitochondrial proteins.
1999,
Pubmed
,
Xenbase
Lakshmipathy,
Mitochondrial DNA ligase III function is independent of Xrcc1.
2000,
Pubmed
,
Xenbase
Le Chalony,
Partial complementation of a DNA ligase I deficiency by DNA ligase III and its impact on cell survival and telomere stability in mammalian cells.
2012,
Pubmed
,
Xenbase
Ohi,
Negative Staining and Image Classification - Powerful Tools in Modern Electron Microscopy.
2004,
Pubmed
Scarff,
Variations on Negative Stain Electron Microscopy Methods: Tools for Tackling Challenging Systems.
2018,
Pubmed
Schneidman-Duhovny,
FoXS, FoXSDock and MultiFoXS: Single-state and multi-state structural modeling of proteins and their complexes based on SAXS profiles.
2016,
Pubmed
Simsek,
Crucial role for DNA ligase III in mitochondria but not in Xrcc1-dependent repair.
2011,
Pubmed
,
Xenbase
Slotboom,
Static light scattering to characterize membrane proteins in detergent solution.
2008,
Pubmed
Sverzhinsky,
Coordinated rearrangements between cytoplasmic and periplasmic domains of the membrane protein complex ExbB-ExbD of Escherichia coli.
2014,
Pubmed
Vijayachandran,
Robots, pipelines, polyproteins: enabling multiprotein expression in prokaryotic and eukaryotic cells.
2011,
Pubmed
Wen,
Size-exclusion chromatography with on-line light-scattering, absorbance, and refractive index detectors for studying proteins and their interactions.
1996,
Pubmed