{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,12]],"date-time":"2026-04-12T14:23:36Z","timestamp":1776003816915,"version":"3.50.1"},"reference-count":42,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2016,1,1]],"date-time":"2016-01-01T00:00:00Z","timestamp":1451606400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2016,1,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Hypomorphic mutations in the nonhomologous end-joining (NHEJ) DNA repair protein DNA ligase IV (LIG4) lead to immunodeficiency with varying severity. In this study, using a murine knock-in model, we investigated the mechanisms underlying abnormalities in class switch recombination (CSR) associated with the human homozygous Lig4 R278H mutation. Previously, we found that despite the near absence of Lig4 end-ligation activity and severely reduced mature B cell numbers, Lig4R278H\/R278H (Lig4R\/R) mice exhibit only a partial CSR block, producing near normal IgG1 and IgE but substantially reduced IgG3, IgG2b, and IgA serum levels. In this study, to address the cause of these abnormalities, we assayed CSR in Lig4R\/R B cells generated via preassembled IgH and IgK V region exons (HL). This revealed that Lig4R278H protein levels while intact exhibited a higher turnover rate during activation of switching to IgG3 and IgG2b, as well as delays in CSR kinetics associated with defective proliferation during activation of switching to IgG1 and IgE. Activated Lig4R\/RHL B cells consistently accumulated high frequencies of activation-induced cytidine deaminase\u2013dependent IgH locus chromosomal breaks and translocations and were more prone to apoptosis, effects that appeared to be p53-independent, as p53 deficiency did not markedly influence these events. Importantly, NHEJ instead of alternative end-joining (A-EJ) was revealed as the predominant mechanism catalyzing robust CSR. Defective CSR was linked to failed NHEJ and residual A-EJ access to unrepaired double-strand breaks. These data firmly demonstrate that Lig4R278H activity renders NHEJ to be more error-prone, and they predict increased error-prone NHEJ activity and A-EJ suppression as the cause of the defective B lymphopoiesis in Lig4 patients.<\/jats:p>","DOI":"10.4049\/jimmunol.1403099","type":"journal-article","created":{"date-parts":[[2015,11,26]],"date-time":"2015-11-26T03:04:22Z","timestamp":1448507062000},"page":"244-255","source":"Crossref","is-referenced-by-count":7,"title":["The DNA Ligase IV Syndrome R278H Mutation Impairs B Lymphopoiesis via Error-Prone Nonhomologous End-Joining"],"prefix":"10.1093","volume":"196","author":[{"given":"Jihye","family":"Park","sequence":"first","affiliation":[{"name":"Department of Pathology, Beth Israel Deaconess Medical Center , Boston, MA 02215"},{"name":"Broad Institute of MIT and Harvard , Cambridge, MA 02142"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert S","family":"Welner","sequence":"additional","affiliation":[{"name":"Harvard Stem Cell Institute, Harvard Medical School , Boston, MA 02115"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mei-Yee","family":"Chan","sequence":"additional","affiliation":[{"name":"Department of Pathology, Beth Israel Deaconess Medical Center , Boston, MA 02215"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Logan","family":"Troppito","sequence":"additional","affiliation":[{"name":"Department of Pathology, Beth Israel Deaconess Medical Center , Boston, MA 02215"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Philipp B","family":"Staber","sequence":"additional","affiliation":[{"name":"Harvard Stem Cell Institute, Harvard Medical School , Boston, MA 02115"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel G","family":"Tenen","sequence":"additional","affiliation":[{"name":"Harvard Stem Cell Institute, Harvard Medical School , Boston, MA 02115"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Catherine T","family":"Yan","sequence":"additional","affiliation":[{"name":"Department of Pathology, Beth Israel Deaconess Medical Center , Boston, MA 02215"},{"name":"Broad Institute of MIT and Harvard , Cambridge, MA 02142"},{"name":"Harvard Stem Cell Institute, Harvard Medical School , Boston, MA 02115"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2016,1,1]]},"reference":[{"key":"2025030702112235200_r1","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/S0065-2776(04)86002-4","article-title":"Mechanism and control of V(D)J recombination versus class switch recombination: similarities and differences","volume":"86","author":"Dudley","year":"2005","journal-title":"Adv. Immunol."},{"key":"2025030702112235200_r2","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/S0065-2776(06)94006-1","article-title":"Evolution of the immunoglobulin heavy chain class switch recombination mechanism","volume":"94","author":"Chaudhuri","year":"2007","journal-title":"Adv. Immunol."},{"key":"2025030702112235200_r3","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1038\/nature06020","article-title":"IgH class switching and translocations use a robust non-classical end-joining pathway","volume":"449","author":"Yan","year":"2007","journal-title":"Nature"},{"key":"2025030702112235200_r4","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1038\/24172","article-title":"Late embryonic lethality and impaired V(D)J recombination in mice lacking DNA ligase IV","volume":"396","author":"Frank","year":"1998","journal-title":"Nature"},{"key":"2025030702112235200_r5","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1259\/0007-1285-63-752-624","article-title":"An instance of clinical radiation morbidity and cellular radiosensitivity, not associated with ataxia-telangiectasia","volume":"63","author":"Plowman","year":"1990","journal-title":"Br. J. Radiol."},{"key":"2025030702112235200_r6","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1016\/S0960-9822(99)80311-X","article-title":"Identification of a defect in DNA ligase IV in a radiosensitive leukaemia patient","volume":"9","author":"Riballo","year":"1999","journal-title":"Curr. Biol."},{"key":"2025030702112235200_r7","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1002\/ajmg.a.30869","article-title":"A patient with mutations in DNA ligase IV: clinical features and overlap with Nijmegen breakage syndrome","volume":"137A","author":"Ben-Omran","year":"2005","journal-title":"Am. J. Med. Genet. A."},{"key":"2025030702112235200_r8","doi-asserted-by":"crossref","first-page":"742","DOI":"10.1002\/ajmg.a.31644","article-title":"Epstein-Barr virus-associated B-cell lymphoma in a patient with DNA ligase IV (LIG4) syndrome","volume":"143A","author":"Toita","year":"2007","journal-title":"Am. J. Med. Genet. A."},{"key":"2025030702112235200_r9","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1016\/S1097-2765(01)00408-7","article-title":"DNA ligase IV mutations identified in patients exhibiting developmental delay and immunodeficiency","volume":"8","author":"O\u2019Driscoll","year":"2001","journal-title":"Mol. Cell"},{"key":"2025030702112235200_r10","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1002\/pbc.22031","article-title":"A novel mutation in a family with DNA ligase IV deficiency syndrome","volume":"53","author":"Unal","year":"2009","journal-title":"Pediatr. Blood Cancer"},{"key":"2025030702112235200_r11","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1002\/humu.22461","article-title":"Extreme growth failure is a common presentation of ligase IV deficiency","volume":"35","author":"Murray","year":"2014","journal-title":"Hum. Mutat."},{"key":"2025030702112235200_r12","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.1016\/j.jaci.2008.08.031","article-title":"Omenn syndrome is associated with mutations in DNA ligase IV","volume":"122","author":"Grunebaum","year":"2008","journal-title":"J. Allergy Clin. Immunol."},{"key":"2025030702112235200_r13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/BF00438824","article-title":"Studies of malformation syndromes of man XXIV B: the Dubowitz syndrome. Further observations","volume":"116","author":"Opitz","year":"1973","journal-title":"Z. Kinderheilkd."},{"key":"2025030702112235200_r14","doi-asserted-by":"crossref","first-page":"e54389","DOI":"10.1371\/journal.pone.0054389","article-title":"Identification of the DNA repair defects in a case of Dubowitz syndrome","volume":"8","author":"Yue","year":"2013","journal-title":"PLoS One"},{"key":"2025030702112235200_r15","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1172\/JCI26121","article-title":"A new type of radiosensitive T\u2212B\u2212NK+ severe combined immunodeficiency caused by a LIG4 mutation","volume":"116","author":"van der Burg","year":"2006","journal-title":"J. Clin. Invest."},{"key":"2025030702112235200_r16","doi-asserted-by":"crossref","first-page":"1611","DOI":"10.1002\/humu.22436","article-title":"Clinical spectrum of LIG4 deficiency is broadened with severe dysmaturity, primordial dwarfism, and neurological abnormalities","volume":"34","author":"IJspeert","year":"2013","journal-title":"Hum. Mutat."},{"key":"2025030702112235200_r17","doi-asserted-by":"crossref","first-page":"5060","DOI":"10.4049\/jimmunol.176.8.5060","article-title":"A severe form of human combined immunodeficiency due to mutations in DNA ligase IV","volume":"176","author":"Enders","year":"2006","journal-title":"J. Immunol."},{"key":"2025030702112235200_r18","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1084\/jem.20040772","article-title":"Impact of DNA ligase IV on nonhomologous end joining pathways during class switch recombination in human cells","volume":"201","author":"Pan-Hammarstr\u00f6m","year":"2005","journal-title":"J. Exp. Med."},{"key":"2025030702112235200_r19","doi-asserted-by":"crossref","first-page":"31124","DOI":"10.1074\/jbc.M103866200","article-title":"Cellular and biochemical impact of a mutation in DNA ligase IV conferring clinical radiosensitivity","volume":"276","author":"Riballo","year":"2001","journal-title":"J. Biol. Chem."},{"key":"2025030702112235200_r20","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/S0921-8777(98)00063-9","article-title":"Absence of DNA ligase IV protein in XR-1 cells: evidence for stabilization by XRCC4","volume":"433","author":"Bryans","year":"1999","journal-title":"Mutat. Res."},{"key":"2025030702112235200_r21","doi-asserted-by":"crossref","first-page":"3024","DOI":"10.1073\/pnas.0914865107","article-title":"Homozygous DNA ligase IV R278H mutation in mice leads to leaky SCID and represents a model for human LIG4 syndrome","volume":"107","author":"Rucci","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025030702112235200_r22","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1016\/S1097-2765(00)80264-6","article-title":"DNA ligase IV deficiency in mice leads to defective neurogenesis and embryonic lethality via the p53 pathway","volume":"5","author":"Frank","year":"2000","journal-title":"Mol. Cell"},{"key":"2025030702112235200_r23","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/S1074-7613(00)80325-8","article-title":"B cell development under the condition of allelic inclusion","volume":"6","author":"Sonoda","year":"1997","journal-title":"Immunity"},{"key":"2025030702112235200_r24","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1016\/S1074-7613(00)80395-7","article-title":"Receptor editing in a transgenic mouse model: site, efficiency, and role in B cell tolerance and antibody diversification","volume":"7","author":"Pelanda","year":"1997","journal-title":"Immunity"},{"key":"2025030702112235200_r25","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1038\/356215a0","article-title":"Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours","volume":"356","author":"Donehower","year":"1992","journal-title":"Nature"},{"key":"2025030702112235200_r26","doi-asserted-by":"crossref","first-page":"3034","DOI":"10.1073\/pnas.0915067107","article-title":"Alternative end-joining catalyzes robust IgH locus deletions and translocations in the combined absence of ligase 4 and Ku70","volume":"107","author":"Boboila","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025030702112235200_r27","doi-asserted-by":"crossref","first-page":"1541","DOI":"10.1158\/1535-7163.MCT-05-0193","article-title":"Radiosensitization of MDA-MB-231 breast tumor cells by adenovirus-mediated overexpression of a fragment of the XRCC4 protein","volume":"4","author":"Jones","year":"2005","journal-title":"Mol. Cancer Ther."},{"key":"2025030702112235200_r28","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/j.cell.2007.06.016","article-title":"ATM prevents the persistence and propagation of chromosome breaks in lymphocytes","volume":"130","author":"Call\u00e9n","year":"2007","journal-title":"Cell"},{"key":"2025030702112235200_r29","doi-asserted-by":"crossref","first-page":"1696","DOI":"10.1172\/JCI32743","article-title":"Impaired lymphocyte development and antibody class switching and increased malignancy in a murine model of DNA ligase IV syndrome","volume":"119","author":"Nijnik","year":"2009","journal-title":"J. Clin. Invest."},{"key":"2025030702112235200_r30","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.dnarep.2008.08.011","article-title":"Mammalian SWI\/SNF chromatin remodeling complexes are required to prevent apoptosis after DNA damage","volume":"8","author":"Park","year":"2009","journal-title":"DNA Repair (Amst.)"},{"key":"2025030702112235200_r31","doi-asserted-by":"crossref","first-page":"14553","DOI":"10.1073\/pnas.241525998","article-title":"Switch junction sequences in PMS2-deficient mice reveal a microhomology-mediated mechanism of Ig class switch recombination","volume":"98","author":"Ehrenstein","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025030702112235200_r32","doi-asserted-by":"crossref","first-page":"2473","DOI":"10.1073\/pnas.1121470109","article-title":"Robust chromosomal DNA repair via alternative end-joining in the absence of X-ray repair cross-complementing protein 1 (XRCC1)","volume":"109","author":"Boboila","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025030702112235200_r33","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/j.molcel.2006.01.005","article-title":"H2AX prevents DNA breaks from progressing to chromosome breaks and translocations","volume":"21","author":"Franco","year":"2006","journal-title":"Mol. Cell"},{"key":"2025030702112235200_r34","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1084\/jem.20092449","article-title":"Alternative end-joining catalyzes class switch recombination in the absence of both Ku70 and DNA ligase 4","volume":"207","author":"Boboila","year":"2010","journal-title":"J. Exp. Med."},{"key":"2025030702112235200_r35","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1093\/intimm\/10.3.325","article-title":"Ig heavy chain class switching in Rag-deficient mice","volume":"10","author":"Lansford","year":"1998","journal-title":"Int. Immunol."},{"key":"2025030702112235200_r36","doi-asserted-by":"crossref","first-page":"2717","DOI":"10.1073\/pnas.0812304106","article-title":"Integrity of the AID serine-38 phosphorylation site is critical for class switch recombination and somatic hypermutation in mice","volume":"106","author":"Cheng","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025030702112235200_r37","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.immuni.2007.05.018","article-title":"Class switch recombination and somatic hypermutation in early mouse B cells are mediated by B cell and Toll-like receptors","volume":"27","author":"Han","year":"2007","journal-title":"Immunity"},{"key":"2025030702112235200_r38","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1038\/nature08159","article-title":"Mechanisms promoting translocations in editing and switching peripheral B cells","volume":"460","author":"Wang","year":"2009","journal-title":"Nature"},{"key":"2025030702112235200_r39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0065-2776(06)93001-6","article-title":"Class switch recombination: a comparison between mouse and human","volume":"93","author":"Pan-Hammarstr\u00f6m","year":"2007","journal-title":"Adv. Immunol."},{"key":"2025030702112235200_r40","doi-asserted-by":"crossref","first-page":"885","DOI":"10.1038\/ncb1444","article-title":"Ku70 stimulates fusion of dysfunctional telomeres yet protects chromosome ends from homologous recombination","volume":"8","author":"Celli","year":"2006","journal-title":"Nat. Cell Biol."},{"key":"2025030702112235200_r41","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1016\/j.molcel.2014.08.002","article-title":"Chromosomal translocations in human cells are generated by canonical nonhomologous end-joining","volume":"55","author":"Ghezraoui","year":"2014","journal-title":"Mol. Cell"},{"key":"2025030702112235200_r42","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1002\/eji.200535401","article-title":"Severe combined immunodeficiency and microcephaly in siblings with hypomorphic mutations in DNA ligase IV","volume":"36","author":"Buck","year":"2006","journal-title":"Eur. J. Immunol."}],"container-title":["The Journal of Immunology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jimmunol\/article-pdf\/196\/1\/244\/62252802\/1403099.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jimmunol\/article-pdf\/196\/1\/244\/62252802\/1403099.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,7]],"date-time":"2025-03-07T02:13:33Z","timestamp":1741313613000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jimmunol\/article\/196\/1\/244\/7973609"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,1,1]]},"references-count":42,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2016,1,1]]}},"URL":"https:\/\/doi.org\/10.4049\/jimmunol.1403099","relation":{},"ISSN":["0022-1767","1550-6606"],"issn-type":[{"value":"0022-1767","type":"print"},{"value":"1550-6606","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2016,1]]},"published":{"date-parts":[[2016,1,1]]}}}