Organization of kappa light chain genes in germ-line and somatic tissue.

Joho, Rolf H.; Weissman, Irving L.; Early, P.; Cole, Jim; Hood, Leroy

doi:10.1073/pnas.77.2.1106

Cited by 53 publications

(20 citation statements)

References 43 publications

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“…Nucleotide sequences were determined by the Maxam and Gilbert technique (14). High molecular weight DNA was prepared from sperm according to Joho et al (9) and from liver as described by Nottenburg and Weissman (15). ACt) at amino acid 45 rather than the glutamine (CAG) codon found in the sequenced germ-line gene, we could test for the existence ofan M511 germ-line gene by examining whether the Alu I site (A-G-C-T) between amino acids 45 and 46 is lost by any germ-line VK24 genes.…”

Section: Methodsmentioning

confidence: 99%

“…Previously, we characterized the organization of the VKM167 gene family in germ-line DNA by Southern blotting, and we demonstrated that no major changes in the size of the VKM167-related genes occur during embryogenesis, ruling out embryonic episomal insertion of genetic elements (9). In this paper, we describe the isolation of several closely related VK gene clones from a A phage library of mouse sperm DNA by probing with a MOPC 167 (M167) light chain cDNA (pl67kRI).…”

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confidence: 94%

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Somatic diversification is required to generate the V kappa genes of MOPC 511 and MOPC 167 myeloma proteins.

Gershenfeld¹,

Tsukamoto²,

Weissman³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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The immune response to phosphocholine in BALB/c mice involves one group of heavy chain variable region (VH) genes and at least three groups of light chain variable region (VK) genes, represented by the gene products of the myelomas TEPC 15, MOPC 603, and MOPC 167/MOPC 511. The amino acid sequences of BALB/c myeloma c chains MOPC 167 and MOPC 511 are known, and they differ by six amino acids. We have isolated several closely related V region genes of immunoglobulin light chains from a mouse sperm DNA phage library, selecting clones that cross-hybridize with a cDNA plasmid probe encoding the light chain of MOPC 167. We identified six strongly hybridizing clones, representing three separate cloning events. We determined the sequence of the coding and immediate flanking regions of three clones, representing the three separate cloning events, and they proved to be identical. This germ-line sequence encoded the amino acid sequence ofneither MOPC 167 nor MOPC 511, but required four base pair changes to generate the VM167 cDNA sequence and five base pair changes to generate the VcM5ll gene. By Southern hybridization experiments, we demonstrated that neither MOPC 511 nor MOPC 167 germ-line genes exist. We conclude that the VM167 and VKM511 genes are created somatically.An immunoglobulin polypeptide contains two distinct regions, the variable (V) and the constant (C) regions. The variability of amino acid sequences within the V region determines the antigen-binding specificity of the antibody. To explain the origin of antibody diversity, the germline hypothesis proposed that diversity arises from the multiplicity of the V region genes encoded in the germ line, whereas the somatic mutation hypothesis proposed that diversity arises from a few inherited germline genes, which during the life of an individual (ontogeny) undergo somatic variations to create an expanded repertoire of V genes (1-7).From molecular hybridization (and Southern blotting) data gathered by using subgroup-specific VK or VH probes (H, heavy chain), the number of cross-hybridizing germ-line V genes is estimated to range between 4 and 20 genes per subgroup, with 100 VK subgroups (8). However, the question of how much expressed antibody diversity is encoded in the germ-line V genes and how much, if any, is attributable to somatic mechanisms remains unanswered.Previously, we characterized the organization of the VKM167 gene family in germ-line DNA by Southern blotting, and we demonstrated that no major changes in the size of the VKM167-related genes occur during embryogenesis, ruling out embryonic episomal insertion of genetic elements (9). In this paper, we describe the isolation of several closely related VK gene clones from a A phage library of mouse sperm DNA by probing with a MOPC 167 (M167) light chain cDNA (pl67kRI). Because both VKM167 and VKM511 genes belong to the VK24 light chain subgroup, we shall refer to genomic germ-line clones hybridizing to pl67kRI as VK24 genes. We have determined the entire nucleotide sequence of the coding and im...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 94%

Somatic diversification is required to generate the V kappa genes of MOPC 511 and MOPC 167 myeloma proteins.

Gershenfeld¹,

Tsukamoto²,

Weissman³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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“…1 and CTL.5), a thymocyte hybridoma (23C), and livers ofthe appropriate strains ofmice were digested with restriction enzymes, subjected to electrophoresis on agarose gels, blotted to nitrocellulose filters, and hybridized with 32P-labeled DNAs coding for various immunoglobulins, or portions thereof. Liver cells contain immunoglobulin genes in the germ-line context, as in sperm or embryo cells (42,(45)(46)(47).…”

Section: Cells Thymus Derived Lymphocytes (T Cells) Like Antibody-pmentioning

confidence: 99%

Expression of an antigen receptor on T cells does not require recombination at the immunoglobulin JH-C mu locus.

Palladino¹,

Marcu²,

Stavnezer³

1981

Proc. Natl. Acad. Sci. U.S.A.

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Considerable evidence has accumulated suggesting that the antigen receptor(s) on T cells is coded for by genes for the variable (V) region of the immunoglobulin heavy (H) chains. In B cells, a complete gene for the immunoglobulin VH region is formed by somatic recombination of VH and joining region heavy chain (JH) gene segments [through an intermediate diversity (D) region gene segment]. In an attempt to determine whether a complete immunoglobulin VH region is expressed on T cells that.bear an antigen receptor, we analyzed the restriction map of the JH-CA locus in genomic DNA from two cloned murine cytotoxic T-lymphocyte (CTL) lines specific for the x-ray-induced leukemia RLCT. We found no rearrangement of the JH-CM locus in the CM lines, indicating that the T-cell antigen receptor(s) in these CTLs is not coded for by a complete immunoglobulin VH gene formed by joining of VH, (DH), and JH genes. In addition, we determined that C,, genes on both chromosomes were present and.that there. was no rearrangement of the C., C.K or A chain genes in these CTL. cells.Thymus derived lymphocytes (T cells), like antibody-producing cells (B cells), show an exquisite specificity in their capacity to recognize antigens. Although it is well documented that membrane immunoglobulins serve as the antigen receptor on B cells (1-3), the nature of the. T-cell receptor(s) remains obscure. Serological and genetic data have suggested that the T-cell receptor(s) is coded for by heavy (H) chain variable region (V) genes (4-15), but whether the receptor(s) is also coded for by H chain constant region (C) genes or light (L) chain V or C region genes has been widely debated (16)(17)(18)(19)(20).In around the JH and C,. genes were altered relative to germ-line DNA. The location ofthe restriction sites that were altered suggested that DNA recombination events had occurred at the JH locus, as would be expected if a VH (D) gene had recombined with a JH gene.To investigate the putative recombination event at the JH-CA locus in T cells that bear an antigen-specific receptor, we hybridized various immunoglobulin cDNA and genomic DNA probes with Southern blots (30) of genomic DNA to examine the restriction maps of the JH-C,, Ca, CK, and A (both Vand C) gene loci in two cloned murine CTL cell lines. Our results indicate that no rearrangement at any of these loci has occurred. We also examined the-immunoglobulin gene loci in a cloned T cell hybridoma line made from thymocytes from a nonimmunized mouse. Again, no rearrangement of the immunoglobulin gene loci was detected.MATERIALS AND METHODS Cell Lines. To produce tumor-specific CTL cell lines, (BALB/ c X C57BL/6)F1 mice were immunized by intradermal injection of 5 X 105 viable BALB/c x-ray-induced leukemia (RLc 1) cells as described (31). Spleen cells from immunized mice, 2 months after the tumor had regressed, were resensitized in vitro with irradiated RLd1 cells for 6 days. These cells were grown in medium containing T-cell growth factor, prepared from CD rat spleen cells according to th...

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“…The J region genes are clustered within a 1.7-kb genomic segment (46,62) and are located 2.4 kb from the CK gene (46,47) on the same EcoRI segment (43). Only one CK exon has been identified (34). At D'Hoostelaere and Huppi, personal communication).…”

mentioning

confidence: 99%

The mouse immunoglobulin kappa light-chain genes are located in early- and late-replicating regions of chromosome 6.

Hatton¹,

Schildkraut²

1990

Mol. Cell. Biol.

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The murine immunoglobulin kappa (K) light-chain multigene family includes the constant region (CK), joining-region genes, and approximately 30 kappa-variable (VK) region families. The entire region occupies an estimated 1,000 to 3,000 kilobases, and some VK families have been linked by recombinant inbred mapping. The CK gene and 14 VK families replicated differently among cell lines of lymphoid and nonlymphoid origin. In nonlymphoid cells, the CK gene replicated earlier than the VK families. A transition from replication during the second third of S phase for the CK gene to later replication during S for VK families was observed. The VK family (VK21) that maps closest to the CK gene, replicated during the first half of the S phase; most of the other VK families replicated during the second half of S, and some replicated during the last quarter of the S phase. In lymphoid cells, the K locus replicated earlier in the pre-B than in the B-cell lines. In one pre-B-cell line, 22D6, the K genes examined replicated at the beginning of the S phase. In the B-cell lines, the EcoRI segment containing the transcribed gene replicated near the beginning of the S phase. Other VKc families replicated within the first two-thirds of S phase. Some linked VK families replicated at similar times. In the B-cell lines, a transition from replication at the beginning of S for the transcribed CK and VK genes and surrounding DNA sequences to later replication for the other VK families was observed. However, in contrast to the non-lymphoid cell lines, the replication of this locus occurred predominantly during the first half of S. The K locus contains both early-and late-replicating genes, and early replication is usually associated with transcriptional activity. The results are discussed with respect to the organization of transcriptionally active chromatin domains. 300 (7, 12, 18, 22, 40, 52, 55, 57). Based on results from the VK21 locus, which determined that the VK genes are separated by about 10 kb (29), the entire K locus could occupy 1,000 to 3,000 kb. A suggested gene order from crossover analysis of congenic and recombinant inbred strains of mice has been produced (5,16,17,22,25,40,61,68). The order of the kappa variable-region genes relative to the centromere and telomere was inferred from these studies (17). The gene order was determined to be centromere; hypodactyly; (IgK-V11, IgK-V24, and IgK-V9-26); (IgK-V9 and IgK-V1); IgK-V12,13; (IgK-V4, IgK-V8, IgK-V1O, IgK-V19); (IgK-V28 and Rn7s-6); IgK-V23; (IgK-V21, IgK-J, and IgK-C); (Ly2, Ly3); waved-1; and telomere (17) Contiguous

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Organization of kappa light chain genes in germ-line and somatic tissue.

Cited by 53 publications

References 43 publications

Somatic diversification is required to generate the V kappa genes of MOPC 511 and MOPC 167 myeloma proteins.

Somatic diversification is required to generate the V kappa genes of MOPC 511 and MOPC 167 myeloma proteins.

Expression of an antigen receptor on T cells does not require recombination at the immunoglobulin JH-C mu locus.

The mouse immunoglobulin kappa light-chain genes are located in early- and late-replicating regions of chromosome 6.

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