{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T11:33:44Z","timestamp":1791286424652,"version":"4.1.0"},"reference-count":52,"publisher":"American Society of Hematology","issue":"8","content-domain":{"domain":["ashpublications.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2001,10,15]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>T-cell receptor (TCR) gene rearrangements are mediated via V(D)J recombination, which is strictly regulated during lymphoid differentiation, most probably through the action of specific transcription factors. Investigated was whether cotransfection ofRAG1 and RAG2 genes in combination with lymphoid transcription factors can induce TCR gene rearrangements in nonlymphoid human cells. Transfection experiments showed that basic helix-loop-helix transcription factors E2A and HEB induce rearrangements in the TCRD locus (D\u03b42-D\u03b43 and V\u03b42-D\u03b43) and TCRG locus (\u03c8 V\u03b37-J\u03b32.3 and V\u03b38-J\u03b32.3). Analysis of these rearrangements and their circular excision products revealed some peculiar characteristics. The V\u03b42-D\u03b43 rearrangements were formed by direct coupling without intermediate D\u03b42 gene segment usage, and most D\u03b42-D\u03b43 recombinations occurred via direct coupling of the respective upstream and downstream recombination signal sequences (RSSs) with deletion of the D\u03b42 and D\u03b43 coding sequences. Subsequently, the E2A\/HEB\u2013induced TCR gene recombination patterns were compared with those in early thymocytes and acute lymphoblastic leukemias of T- and B-lineage origin, and it was found that the TCR rearrangements in the transfectants were early (immature) and not necessarily T-lineage specific. Apparently, some parts of theTCRD (V\u03b42-D\u03b4 region) and TCRG genes are accessible for recombination not only in T cells, but also in early B-cells and even in nonlymphoid cells if the appropriate transcription factors are present. The transfection system described here appeared to be useful for studying the accessibility of immunoglobulin and TCR genes for V(D)J recombination, but might also be applied to study the induction of RSS-mediated chromosome aberrations.<\/jats:p>","DOI":"10.1182\/blood.v98.8.2456","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T12:46:47Z","timestamp":1027774007000},"page":"2456-2465","update-policy":"https:\/\/doi.org\/10.1182\/blood.2019cm0000","source":"Crossref","is-referenced-by-count":59,"title":["Basic helix-loop-helix proteins E2A and HEB induce immature T-cell receptor rearrangements in nonlymphoid cells"],"prefix":"10.1182","volume":"98","author":[{"given":"Anton W.","family":"Langerak","sequence":"first","affiliation":[{"name":"From the Department of Immunology, Erasmus University Rotterdam\/University Hospital Rotterdam, The Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ingrid L. M.","family":"Wolvers-Tettero","sequence":"additional","affiliation":[{"name":"From the Department of Immunology, Erasmus University Rotterdam\/University Hospital Rotterdam, The Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ellen J.","family":"van Gastel-Mol","sequence":"additional","affiliation":[{"name":"From the Department of Immunology, Erasmus University Rotterdam\/University Hospital Rotterdam, The Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monique E. C. M.","family":"Oud","sequence":"additional","affiliation":[{"name":"From the Department of Immunology, Erasmus University Rotterdam\/University Hospital Rotterdam, The Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacques J. M.","family":"van Dongen","sequence":"additional","affiliation":[{"name":"From the Department of Immunology, Erasmus University Rotterdam\/University Hospital Rotterdam, The Netherlands."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"234","reference":[{"key":"2020021217530467100_B1","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1038\/302575a0","article-title":"Somatic generation of antibody diversity.","volume":"302","author":"Tonegawa","year":"1983","journal-title":"Nature."},{"key":"2020021217530467100_B2","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1016\/0092-8674(89)90760-5","article-title":"The V(D)J recombination activating gene, RAG-1.","volume":"59","author":"Schatz","year":"1989","journal-title":"Cell."},{"key":"2020021217530467100_B3","doi-asserted-by":"crossref","first-page":"1517","DOI":"10.1126\/science.2360047","article-title":"RAG-1 and RAG-2, adjacent genes that synergistically activate V(D)J recombination.","volume":"248","author":"Oettinger","year":"1990","journal-title":"Science."},{"key":"2020021217530467100_B4","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1016\/0092-8674(95)90012-8","article-title":"Initiation of V(D)J recombination in a cell-free system.","volume":"81","author":"van Gent","year":"1995","journal-title":"Cell."},{"key":"2020021217530467100_B5","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1146\/annurev.immunol.14.1.459","article-title":"Accessibility control of antigen-receptor variable-region gene assembly: role of cis-acting elements.","volume":"14","author":"Sleckman","year":"1996","journal-title":"Annu Rev Immunol."},{"key":"2020021217530467100_B6","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1038\/sj.leu.2401277","article-title":"Cross-lineage T-cell receptor gene rearrangements occur in more than ninety percent of childhood precursor-B-acute lymphoblastic leukemias: alternative PCR targets for detection of minimal residual disease.","volume":"13","author":"Szczepanski","year":"1999","journal-title":"Leukemia."},{"key":"2020021217530467100_B7","doi-asserted-by":"crossref","first-page":"4079","DOI":"10.1182\/blood.V93.12.4079","article-title":"Ig heavy chain gene rearrangements in T-cell acute lymphoblastic leukemia exhibit predominant DH6\u201319 and DH7\u201327 gene usage, can result in complete V-D-J rearrangements, and are rare in T-cell receptor alpha beta lineage.","volume":"93","author":"Szczepanski","year":"1999","journal-title":"Blood."},{"key":"2020021217530467100_B8","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/0092-8674(85)90141-2","article-title":"Developmentally controlled and tissue-specific expression of unrearranged VH gene segments.","volume":"40","author":"Yancopoulos","year":"1985","journal-title":"Cell."},{"key":"2020021217530467100_B9","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/0009-8981(91)90247-A","article-title":"Analysis of immunoglobulin and T-cell receptor genes, II: possibilities and limitations in the diagnosis and management of lymphoproliferative diseases and related disorders.","volume":"198","author":"Van Dongen","year":"1991","journal-title":"Clin Chim Acta."},{"key":"2020021217530467100_B10","doi-asserted-by":"crossref","first-page":"4120","DOI":"10.4049\/jimmunol.156.11.4120","article-title":"Complete TCR-delta rearrangements and partial (D-J) recombination of the TCR-beta locus in CD34+7+ precursors from human cord blood.","volume":"156","author":"Ktorza","year":"1996","journal-title":"J Immunol."},{"key":"2020021217530467100_B11","doi-asserted-by":"crossref","first-page":"3033","DOI":"10.1182\/blood.V93.9.3033","article-title":"TCR gene rearrangements and expression of the pre-T cell receptor complex during human T-cell differentiation.","volume":"93","author":"Blom","year":"1999","journal-title":"Blood."},{"key":"2020021217530467100_B12","doi-asserted-by":"crossref","first-page":"1209","DOI":"10.1002\/j.1460-2075.1984.tb01955.x","article-title":"Ordered rearrangement of immunoglobulin heavy chain variable region segments.","volume":"3","author":"Alt","year":"1984","journal-title":"EMBO J."},{"key":"2020021217530467100_B13","doi-asserted-by":"crossref","first-page":"943","DOI":"10.1126\/science.2658058","article-title":"Developmental biology of T cell receptors.","volume":"244","author":"Strominger","year":"1989","journal-title":"Science."},{"key":"2020021217530467100_B14","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1038\/314103a0","article-title":"Developmental regulation of T-cell receptor gene expression.","volume":"314","author":"Raulet","year":"1985","journal-title":"Nature."},{"key":"2020021217530467100_B15","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1084\/jem.189.2.289","article-title":"Positive and negative regulation of V(D)J recombination by the E2A proteins.","volume":"189","author":"Bain","year":"1999","journal-title":"J Exp Med."},{"key":"2020021217530467100_B16","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1016\/S0092-8674(00)81272-6","article-title":"Cell type-specific chromatin structure determines the targeting of V(D)J recombinase activity in vitro.","volume":"85","author":"Stanhope-Baker","year":"1996","journal-title":"Cell."},{"key":"2020021217530467100_B17","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1016\/0092-8674(89)90682-X","article-title":"A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD, and myc proteins.","volume":"56","author":"Murre","year":"1989","journal-title":"Cell."},{"key":"2020021217530467100_B18","doi-asserted-by":"crossref","first-page":"3522","DOI":"10.1128\/MCB.13.6.3522","article-title":"E2A and E2\u20132 are subunits of B-cell-specific E2-box DNA-binding proteins.","volume":"13","author":"Bain","year":"1993","journal-title":"Mol Cell Biol."},{"key":"2020021217530467100_B19","doi-asserted-by":"crossref","first-page":"4518","DOI":"10.1128\/MCB.15.8.4518","article-title":"B-cell-specific DNA binding by an E47 homodimer.","volume":"15","author":"Shen","year":"1995","journal-title":"Mol Cell Biol."},{"key":"2020021217530467100_B20","doi-asserted-by":"crossref","first-page":"885","DOI":"10.1016\/0092-8674(94)90077-9","article-title":"E2A proteins are required for proper B cell development and initiation of immunoglobulin gene rearrangements [comment appears in Cell. 1994;79:751-753].","volume":"79","author":"Bain","year":"1994","journal-title":"Cell."},{"key":"2020021217530467100_B21","doi-asserted-by":"crossref","first-page":"875","DOI":"10.1016\/0092-8674(94)90076-0","article-title":"The helix-loop-helix gene E2A is required for B cell formation.","volume":"79","author":"Zhuang","year":"1994","journal-title":"Cell."},{"key":"2020021217530467100_B22","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1101\/gad.5.8.1367","article-title":"Helix-loop-helix transcription factor E47 activates germ-line immunoglobulin heavy-chain gene transcription and rearrangement in a pre-T-cell line.","volume":"5","author":"Schlissel","year":"1991","journal-title":"Genes Dev."},{"key":"2020021217530467100_B23","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/S1097-2765(00)80429-3","article-title":"E2A and EBF act in synergy with the V(D)J recombinase to generate a diverse immunoglobulin repertoire in nonlymphoid cells.","volume":"5","author":"Romanow","year":"2000","journal-title":"Mol Cell."},{"key":"2020021217530467100_B24","doi-asserted-by":"crossref","first-page":"2898","DOI":"10.1128\/MCB.16.6.2898","article-title":"B-lymphocyte development is regulated by the combined dosage of three basic helix-loop-helix genes, E2A, E2\u20132, and HEB.","volume":"16","author":"Zhuang","year":"1996","journal-title":"Mol Cell Biol."},{"key":"2020021217530467100_B25","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1016\/0092-8674(91)90057-6","article-title":"Translocations, master genes, and differences between the origins of acute and chronic leukemias.","volume":"67","author":"Rabbitts","year":"1991","journal-title":"Cell."},{"key":"2020021217530467100_B26","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1146\/annurev.iy.10.040192.002043","article-title":"V(D)J recombination: molecular biology and regulation.","volume":"10","author":"Schatz","year":"1992","journal-title":"Annu Rev Immunol."},{"key":"2020021217530467100_B27","first-page":"1","article-title":"Chromosomal translocations in lymphoid neoplasia: a reappraisal of the recombinase model.","volume":"2","author":"Tycko","year":"1990","journal-title":"Cancer Cells."},{"key":"2020021217530467100_B28","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1016\/0952-7915(95)80074-3","article-title":"The role of chromosome translocations in T cell acute leukemia.","volume":"7","author":"Hwang","year":"1995","journal-title":"Curr Opin Immunol."},{"key":"2020021217530467100_B29","doi-asserted-by":"crossref","first-page":"8392","DOI":"10.1073\/pnas.90.18.8392","article-title":"Production of high-titer helper-free retroviruses by transient transfection.","volume":"90","author":"Pear","year":"1993","journal-title":"Proc Natl Acad Sci U S A."},{"key":"2020021217530467100_B30","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1084\/jem.188.4.699","article-title":"Induction of early B cell factor (EBF) and multiple B lineage genes by the basic helix-loop-helix transcription factor E12.","volume":"188","author":"Kee","year":"1998","journal-title":"J Exp Med."},{"key":"2020021217530467100_B31","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1128\/MCB.8.1.466","article-title":"SR alpha promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat.","volume":"8","author":"Takebe","year":"1988","journal-title":"Mol Cell Biol."},{"key":"2020021217530467100_B32","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/S1074-7613(00)80506-3","article-title":"Complementation of V(D)J recombination deficiency in RAG-1(\u2212\/\u2212) B cells reveals a requirement for novel elements in the N-terminus of RAG-1.","volume":"7","author":"Roman","year":"1997","journal-title":"Immunity."},{"key":"2020021217530467100_B33","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1038\/sj.leu.2401245","article-title":"Primers and protocols for standardized detection of minimal residual disease in acute lymphoblastic leukemia using immunoglobulin and T cell receptor gene rearrangements and TAL1 deletions as PCR targets: report of the BIOMED-1 CONCERTED ACTION: investigation of minimal residual disease in acute leukemia.","volume":"13","author":"Pongers-Willemse","year":"1999","journal-title":"Leukemia."},{"key":"2020021217530467100_B34","doi-asserted-by":"crossref","first-page":"3930","DOI":"10.1182\/blood.V86.10.3930.bloodjournal86103930","article-title":"Analysis of rearranged T-cell receptor beta-chain genes by polymerase chain reaction (PCR) DNA sequencing and automated high resolution PCR fragment analysis.","volume":"86","author":"Kneba","year":"1995","journal-title":"Blood."},{"key":"2020021217530467100_B35","doi-asserted-by":"crossref","first-page":"4341","DOI":"10.4049\/jimmunol.159.9.4341","article-title":"Human T cell leukemias with continuous V(D)J recombinase activity for TCR-delta gene deletion.","volume":"159","author":"Breit","year":"1997","journal-title":"J Immunol."},{"key":"2020021217530467100_B36","doi-asserted-by":"crossref","first-page":"2192","DOI":"10.1038\/sj.leu.2400887","article-title":"Heteroduplex PCR analysis of rearranged T cell receptor genes for clonality assessment in suspect T cell proliferations.","volume":"11","author":"Langerak","year":"1997","journal-title":"Leukemia."},{"key":"2020021217530467100_B37","doi-asserted-by":"crossref","first-page":"2006","DOI":"10.1038\/sj.leu.2401246","article-title":"Real-time quantitative PCR for the detection of minimal residual disease in acute lymphoblastic leukemia using junctional regions specific TaqMan probes.","volume":"12","author":"Pongers-Willemse","year":"1998","journal-title":"Leukemia."},{"key":"2020021217530467100_B38","doi-asserted-by":"crossref","first-page":"1419","DOI":"10.1038\/sj.leu.2401831","article-title":"Improved assessment of minimal residual disease in B cell malignancies using fluorogenic consensus probes for real-time quantitative PCR.","volume":"14","author":"Bruggemann","year":"2000","journal-title":"Leukemia."},{"key":"2020021217530467100_B39","doi-asserted-by":"crossref","first-page":"1426","DOI":"10.1038\/sj.leu.2401801","article-title":"Application of germline IGH probes in real-time quantitative PCR for the detection of minimal residual disease in acute lymphoblastic leukemia.","volume":"14","author":"Verhagen","year":"2000","journal-title":"Leukemia."},{"key":"2020021217530467100_B40","first-page":"1076","article-title":"Extensive junctional diversity of \u03b3\u03b4 T-cell receptors expressed by T-cell acute lymphoblastic leukemias: implications for the detection of minimal residual disease.","volume":"5","author":"Breit","year":"1991","journal-title":"Leukemia."},{"key":"2020021217530467100_B41","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1038\/sj.leu.2401276","article-title":"Immunophenotypic and immunogenotypic characteristics of TCR\u03b3\u03b4+ T cell acute lymphoblastic leukemia.","volume":"13","author":"Langerak","year":"1999","journal-title":"Leukemia."},{"key":"2020021217530467100_B42","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1002\/1097-0320(20000801)40:4<336::AID-CYTO9>3.0.CO;2-0","article-title":"Flow cytometric analysis of the Vbeta repertoire in healthy controls.","volume":"40","author":"Van den Beemd","year":"2000","journal-title":"Cytometry."},{"key":"2020021217530467100_B43","doi-asserted-by":"crossref","first-page":"745","DOI":"10.4049\/jimmunol.145.2.745","article-title":"Novel recombinations of the IG kappa-locus that result in allelic exclusion.","volume":"145","author":"Feddersen","year":"1990","journal-title":"J Immunol."},{"key":"2020021217530467100_B44","doi-asserted-by":"crossref","first-page":"3063","DOI":"10.1182\/blood.V82.10.3063.3063","article-title":"Southern blot patterns, frequencies and junctional diversity of T-cell receptor-\u03b4 gene rearrangements in acute lymphoblastic leukemia.","volume":"82","author":"Breit","year":"1993","journal-title":"Blood."},{"key":"2020021217530467100_B45","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1038\/sj.leu.2401427","article-title":"Detection of T cell receptor beta (TCRB) gene rearrangement patterns in T cell malignancies by Southern blot analysis.","volume":"13","author":"Langerak","year":"1999","journal-title":"Leukemia."},{"key":"2020021217530467100_B46","doi-asserted-by":"crossref","first-page":"1620","DOI":"10.1038\/sj.leu.2401540","article-title":"Easy detection of all T cell receptor gamma (TCRG) gene rearrangements by Southern blot analysis: recommendations for optimal results.","volume":"13","author":"Moreau","year":"1999","journal-title":"Leukemia."},{"key":"2020021217530467100_B47","doi-asserted-by":"crossref","first-page":"1208","DOI":"10.1038\/sj.leu.2401765","article-title":"T cell receptor gamma (TCRG) gene rearrangements in T cell acute lymphoblastic leukemia reflect \u201cend-stage\u201d recombinations: implications for minimal residual disease monitoring.","volume":"14","author":"Szczepanski","year":"2000","journal-title":"Leukemia."},{"key":"2020021217530467100_B48","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1182\/blood.V77.1.141.141","article-title":"T-cell receptor delta gene recombination in common acute lymphoblastic leukemia: preferential usage of V delta 2 and frequent involvement of the J alpha cluster.","volume":"77","author":"Yokota","year":"1991","journal-title":"Blood."},{"key":"2020021217530467100_B49","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1126\/science.2955517","article-title":"Structurally divergent human T cell receptor \u03b3 proteins encoded by distinct C\u03b3 genes.","volume":"237","author":"Krangel","year":"1987","journal-title":"Science."},{"key":"2020021217530467100_B50","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/S1097-2765(00)80188-4","article-title":"A conserved motif present in a class of helix-loop-helix proteins activates transcription by direct recruitment of the SAGA complex.","volume":"4","author":"Massari","year":"1999","journal-title":"Mol Cell."},{"key":"2020021217530467100_B51","doi-asserted-by":"crossref","first-page":"1461","DOI":"10.1002\/j.1460-2075.1985.tb03803.x","article-title":"The chromosomal location of T-cell receptor genes and a T-cell rearranging gene: possible correlation with specific translocations in human T-cell leukemia.","volume":"4","author":"Rabbitts","year":"1985","journal-title":"EMBO J."},{"key":"2020021217530467100_B52","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1084\/jem.177.4.965","article-title":"Site-specific deletions involving the tal-1 and sil genes are restricted to cells of the T cell receptor alpha\/beta lineage: T cell receptor delta gene deletion mechanism affects multiple genes.","volume":"177","author":"Breit","year":"1993","journal-title":"J Exp Med."}],"container-title":["Blood"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/ashpublications.org\/blood\/article-pdf\/98\/8\/2456\/1677497\/h8200102456.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/ashpublications.org\/blood\/article-pdf\/98\/8\/2456\/1677497\/h8200102456.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,4,23]],"date-time":"2023-04-23T20:10:58Z","timestamp":1682280658000},"score":1,"resource":{"primary":{"URL":"https:\/\/ashpublications.org\/blood\/article\/98\/8\/2456\/53272\/Basic-helixloophelix-proteins-E2A-and-HEB-induce"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2001,10,15]]},"references-count":52,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2001,10,15]]}},"URL":"https:\/\/doi.org\/10.1182\/blood.v98.8.2456","relation":{},"ISSN":["1528-0020","0006-4971"],"issn-type":[{"value":"1528-0020","type":"electronic"},{"value":"0006-4971","type":"print"}],"subject":[],"published":{"date-parts":[[2001,10,15]]}}}