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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...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2009
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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. |
format | Text |
id | pubmed-2774322 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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