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RAD51 paralogs promote homology-directed repair at diversifying immunoglobulin V regions

BACKGROUND: Gene conversion depends upon the same factors that carry out more general process of homologous recombination, including homologous gene targeting and recombinational repair. Among these are the RAD51 paralogs, conserved factors related to the key recombination factor, RAD51. In chicken...

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Autores principales: Ordinario, Ellen C, Yabuki, Munehisa, Handa, Priya, Cummings, W Jason, Maizels, Nancy
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2774322/
https://www.ncbi.nlm.nih.gov/pubmed/19863810
http://dx.doi.org/10.1186/1471-2199-10-98
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author Ordinario, Ellen C
Yabuki, Munehisa
Handa, Priya
Cummings, W Jason
Maizels, Nancy
author_facet Ordinario, Ellen C
Yabuki, Munehisa
Handa, Priya
Cummings, W Jason
Maizels, Nancy
author_sort Ordinario, Ellen C
collection PubMed
description BACKGROUND: Gene conversion depends upon the same factors that carry out more general process of homologous recombination, including homologous gene targeting and recombinational repair. Among these are the RAD51 paralogs, conserved factors related to the key recombination factor, RAD51. In chicken and other fowl, gene conversion (templated mutation) diversifies immunoglobulin variable region sequences. This allows gene conversion and recombinational repair to be studied using the chicken DT40 B cell line, which carries out constitutive gene conversion and provides a robust and physiological model for homology-directed repair in vertebrate cells. RESULTS: We show that DT40 contains constitutive nuclear foci of the repair factors RAD51D and XRCC2, consistent with activated homologous recombination. Single-cell imaging of a DT40 derivative in which the rearranged and diversifying immunoglobulin λ(R )light chain gene is tagged with polymerized lactose operator, DT40 PolyLacO-λ(R), showed that RAD51D and XRCC2 localize to the diversifying λ(R )gene. Colocalizations correlate both functionally and physically with active immunoglobulin gene conversion. Ectopic expression of either RAD51D or XRCC2 accelerated the clonal rate of gene conversion, and conversion tracts were significantly longer in RAD51D than XRCC2 transfectants. CONCLUSION: These results demonstrate direct functions of RAD51D and XRCC2 in immunoglobulin gene conversion, and also suggest that modulation of levels of repair factors may be a useful strategy to promote gene correction in other cell types.
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spelling pubmed-27743222009-11-07 RAD51 paralogs promote homology-directed repair at diversifying immunoglobulin V regions Ordinario, Ellen C Yabuki, Munehisa Handa, Priya Cummings, W Jason Maizels, Nancy BMC Mol Biol Research Article BACKGROUND: Gene conversion depends upon the same factors that carry out more general process of homologous recombination, including homologous gene targeting and recombinational repair. Among these are the RAD51 paralogs, conserved factors related to the key recombination factor, RAD51. In chicken and other fowl, gene conversion (templated mutation) diversifies immunoglobulin variable region sequences. This allows gene conversion and recombinational repair to be studied using the chicken DT40 B cell line, which carries out constitutive gene conversion and provides a robust and physiological model for homology-directed repair in vertebrate cells. RESULTS: We show that DT40 contains constitutive nuclear foci of the repair factors RAD51D and XRCC2, consistent with activated homologous recombination. Single-cell imaging of a DT40 derivative in which the rearranged and diversifying immunoglobulin λ(R )light chain gene is tagged with polymerized lactose operator, DT40 PolyLacO-λ(R), showed that RAD51D and XRCC2 localize to the diversifying λ(R )gene. Colocalizations correlate both functionally and physically with active immunoglobulin gene conversion. Ectopic expression of either RAD51D or XRCC2 accelerated the clonal rate of gene conversion, and conversion tracts were significantly longer in RAD51D than XRCC2 transfectants. CONCLUSION: These results demonstrate direct functions of RAD51D and XRCC2 in immunoglobulin gene conversion, and also suggest that modulation of levels of repair factors may be a useful strategy to promote gene correction in other cell types. BioMed Central 2009-10-28 /pmc/articles/PMC2774322/ /pubmed/19863810 http://dx.doi.org/10.1186/1471-2199-10-98 Text en Copyright © 2009 Ordinario et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Ordinario, Ellen C
Yabuki, Munehisa
Handa, Priya
Cummings, W Jason
Maizels, Nancy
RAD51 paralogs promote homology-directed repair at diversifying immunoglobulin V regions
title RAD51 paralogs promote homology-directed repair at diversifying immunoglobulin V regions
title_full RAD51 paralogs promote homology-directed repair at diversifying immunoglobulin V regions
title_fullStr RAD51 paralogs promote homology-directed repair at diversifying immunoglobulin V regions
title_full_unstemmed RAD51 paralogs promote homology-directed repair at diversifying immunoglobulin V regions
title_short RAD51 paralogs promote homology-directed repair at diversifying immunoglobulin V regions
title_sort rad51 paralogs promote homology-directed repair at diversifying immunoglobulin v regions
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2774322/
https://www.ncbi.nlm.nih.gov/pubmed/19863810
http://dx.doi.org/10.1186/1471-2199-10-98
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