Insertion of Phosphoglycerine Kinase (PGK)-neo 5' of Jlambda1 Dramatically Enhances VJlambda1 Rearrangement

Overview

Journal J Exp Med

Specialties Allergy & Immunology
General Medicine

Date 2001 Mar 21

PMID 11257137

Citations 7

Authors

T Sun

U STORB

Affiliations

Soon will be listed here.

Abstract

Gene-targeted mice were generated with a loxP-neomycin resistance gene (neo(r)) cassette inserted upstream of the Jlambda1 region and replacement of the glycine 154 codon in the Clambda1 gene with a serine codon. This insertion dramatically increases Vlambda1-Jlambda1 recombination. Jlambda1 germline transcription levels in pre-B cells and thymus cells are also greatly increased, apparently due to the strong housekeeping phosphoglycerine kinase (PGK) promoter driving the neo gene. In contrast, deletion of the neo gene causes a significant decrease in VJlambda1 recombination to levels below those in normal mice. This reduction is due to the loxP site left on the chromosome which reduces the Jlambda1 germline transcription in cis. Thus, the correlation between germline transcription and variable (V), diversity (D), and joining (J) recombination is not just an all or none phenomenon. Rather, the transcription efficiency is directly associated with the recombination efficiency. Furthermore, Jlambda1 and Vlambda1 germline transcription itself is not sufficient to lead to VJ recombination in T cells or early pre-B cells. The findings may suggest that in vivo: (a) locus and cell type-specific transactivators direct the immunoglobulin or T cell receptor loci, respectively, to a "recombination factory" in the nucleus, and (b) transcription complexes deliver V(D)J recombinase to the recombination signal sequences.

Citing Articles

Photoreceptor avascular privilege is shielded by soluble VEGF receptor-1.

Luo L, Uehara H, Zhang X, Das S, Olsen T, Holt D Elife. 2013; 2:e00324.

PMID: 23795287 PMC: 3687373. DOI: 10.7554/eLife.00324.

Enhanced expression of human cDNA by phosphoglycerate kinase promoter-puromycin cassette in the mouse transthyretin locus.

Li Z, Zhao G, Shen J, Araki K, Haruna K, Inoue S Transgenic Res. 2010; 20(1):191-200.

PMID: 20369382 DOI: 10.1007/s11248-010-9389-2.

Cis-regulatory elements and epigenetic changes control genomic rearrangements of the IgH locus.

Perlot T, Alt F Adv Immunol. 2009; 99:1-32.

PMID: 19117530 PMC: 3115588. DOI: 10.1016/S0065-2776(08)00601-9.

The very 5' end and the constant region of Ig genes are spared from somatic mutation because AID does not access these regions.

Longerich S, Tanaka A, Bozek G, Nicolae D, Storb U J Exp Med. 2005; 202(10):1443-54.

PMID: 16301749 PMC: 2212980. DOI: 10.1084/jem.20051604.

IRF-4,8 orchestrate the pre-B-to-B transition in lymphocyte development.

Lu R, Medina K, Lancki D, Singh H Genes Dev. 2003; 17(14):1703-8.

PMID: 12832394 PMC: 196178. DOI: 10.1101/gad.1104803.

References

McMurry M, Krangel M . A role for histone acetylation in the developmental regulation of VDJ recombination. Science. 2000; 287(5452):495-8. DOI: 10.1126/science.287.5452.495. View

Kingston R, Narlikar G . ATP-dependent remodeling and acetylation as regulators of chromatin fluidity. Genes Dev. 1999; 13(18):2339-52. DOI: 10.1101/gad.13.18.2339. View

Miller J, Selsing E, STORB U . Structural alterations in J regions of mouse immunoglobulin lambda genes are associated with differential gene expression. Nature. 1982; 295(5848):428-30. DOI: 10.1038/295428a0. View

Alt F, Rosenberg N, Enea V, Siden E, Baltimore D . Multiple immunoglobulin heavy-chain gene transcripts in Abelson murine leukemia virus-transformed lymphoid cell lines. Mol Cell Biol. 1982; 2(4):386-400. PMC: 369803. DOI: 10.1128/mcb.2.4.386-400.1982. View

Weiss S, Lehmann K, Cohn M . Monoclonal antibodies to murine immunoglobulin isotypes. Hybridoma. 1983; 2(1):49-54. DOI: 10.1089/hyb.1983.2.49. View

Yancopoulos G, Alt F . Developmentally controlled and tissue-specific expression of unrearranged VH gene segments. Cell. 1985; 40(2):271-81. DOI: 10.1016/0092-8674(85)90141-2. View

Landau N, Schatz D, Rosa M, Baltimore D . Increased frequency of N-region insertion in a murine pre-B-cell line infected with a terminal deoxynucleotidyl transferase retroviral expression vector. Mol Cell Biol. 1987; 7(9):3237-43. PMC: 367960. DOI: 10.1128/mcb.7.9.3237-3243.1987. View

Alt F, Blackwell T, Yancopoulos G . Development of the primary antibody repertoire. Science. 1987; 238(4830):1079-87. DOI: 10.1126/science.3317825. View

Miller J, Ogden S, McMullen M, Andres H, STORB U . The order and orientation of mouse lambda-genes explain lambda-rearrangement patterns. J Immunol. 1988; 141(7):2497-502. View

10.

STORB U, Haasch D, Arp B, Sanchez P, Cazenave P, Miller J . Physical linkage of mouse lambda genes by pulsed-field gel electrophoresis suggests that the rearrangement process favors proximate target sequences. Mol Cell Biol. 1989; 9(2):711-8. PMC: 362648. DOI: 10.1128/mcb.9.2.711-718.1989. View

11.

Bogen B . Monoclonal antibodies specific for variable and constant domains of murine lambda chains. Scand J Immunol. 1989; 29(3):273-9. DOI: 10.1111/j.1365-3083.1989.tb01125.x. View

12.

Feeney A . Lack of N regions in fetal and neonatal mouse immunoglobulin V-D-J junctional sequences. J Exp Med. 1990; 172(5):1377-90. PMC: 2188672. DOI: 10.1084/jem.172.5.1377. View

13.

Alessandrini A, Desiderio S . Coordination of immunoglobulin DJH transcription and D-to-JH rearrangement by promoter-enhancer approximation. Mol Cell Biol. 1991; 11(4):2096-107. PMC: 359897. DOI: 10.1128/mcb.11.4.2096-2107.1991. View

14.

Artelt P, Grannemann R, Stocking C, Friel J, Bartsch J, Hauser H . The prokaryotic neomycin-resistance-encoding gene acts as a transcriptional silencer in eukaryotic cells. Gene. 1991; 99(2):249-54. DOI: 10.1016/0378-1119(91)90134-w. View

15.

Hardy R, Carmack C, Shinton S, Kemp J, Hayakawa K . Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow. J Exp Med. 1991; 173(5):1213-25. PMC: 2118850. DOI: 10.1084/jem.173.5.1213. View

16.

Engler P, Haasch D, Pinkert C, Doglio L, Glymour M, Brinster R . A strain-specific modifier on mouse chromosome 4 controls the methylation of independent transgene loci. Cell. 1991; 65(6):939-47. DOI: 10.1016/0092-8674(91)90546-b. View

17.

Ramsden D, Wu G . Mouse kappa light-chain recombination signal sequences mediate recombination more frequently than do those of lambda light chain. Proc Natl Acad Sci U S A. 1991; 88(23):10721-5. PMC: 53002. DOI: 10.1073/pnas.88.23.10721. View

18.

Hsieh C, Lieber M . CpG methylated minichromosomes become inaccessible for V(D)J recombination after undergoing replication. EMBO J. 1992; 11(1):315-25. PMC: 556452. DOI: 10.1002/j.1460-2075.1992.tb05054.x. View

19.

Schatz D, Oettinger M, Schlissel M . V(D)J recombination: molecular biology and regulation. Annu Rev Immunol. 1992; 10:359-83. DOI: 10.1146/annurev.iy.10.040192.002043. View

20.

Engler P, Weng A, STORB U . Influence of CpG methylation and target spacing on V(D)J recombination in a transgenic substrate. Mol Cell Biol. 1993; 13(1):571-7. PMC: 358936. DOI: 10.1128/mcb.13.1.571-577.1993. View