Lambda Locus Research Articles

Analyses of chicken immunoglobulin light chain cDNA clones indicate a few germline V lambda genes and allotypes of the C lambda locus.

cDNA libraries of chicken spleen and Harder gland (a gland enriched with immunocytes) constructed in pBR322 were screened by differential hybridization and by mRNA hybrid-selected translation. Eleven L-chain cDNA clones were identified from which VL probes were prepared and each was annealed with kidney DNA restriction digests. All VL probes revealed the same set of bands, corresponding to about 15 germline VL genes of one subgroup. The nucleotide sequences of six VL clones showed greater than or equal to 85% homology, and the predicted amino acid sequences were identical or nearly identical to the major N-terminal sequence of L-chains in chicken serum. These findings, and the fact that the VL clones were randomly selected from normal lymphoid tissues, strongly indicate that the bulk of chicken L-chains is encoded by a few germline VL genes, probably much less than 15 since many of the VL genes are known to be pseudogenes. Therefore, it is likely that somatic mechanisms operating prior to specific triggering by antigen play a major role in the generation of antibody diversity in chicken. Analysis of the constant region locus (sequencing of CL gene and cDNAs) demonstrate a single CL isotype and suggest the presence of CL allotypes.

The immunoglobulin lambda locus in rat consists of two Cλ genes and a single Vλ gene

The immunoglobin 2 locus of the rat has been studied. Germ-line Vλ and Cλ genes were isolated from recombinant-phage libraries and characterized by nucleotide sequencing. The results showed that the λ locus of the rat contains one single Vλ gene and two Cλ genes, thus representing one of the least complex λ loci so far characterized. The two Cλ genes are separated by a spacer approx. 3 kb long. Two J segments are located at the 5' side of each Cλ gene. One of the Cλ genes (Cλ 1) probably represents a pseudogene, as the Jλ 1 segments have non-functional recombination and splice signals. The organization of the rat λ locus resembles that of mouse, except that only one cluster is present in the rat. Thus since the evolutionary separation of the rat and mouse species ten MYR ( = 10 6 years) ago, either one cluster has been lost from the rat, or duplicated in the mouse.

Human ? light chain locus: Organization and DNA sequences of three genomicJ regions

Evidence for the genomic organization of human lambda light chain joining (J) region gene segments is presented. A mouse J lambda probe was used in Southern hybridizations to localize joining region sequences in a cosmid clone containing the genomic cluster of six human lambda constant (C) region gene segments. The results of these hybridizations suggest the presence of at least one J gene segment upstream from each constant region gene segment. The DNA sequences indicate that the human J lambda 1, J lambda 2, and J lambda 3 gene segments have consensus nonamer and heptamer sequences, proposed to be involved in V-J joining, are capable of encoding the known amino acid sequences for the respective J peptides, and have a sequence which could give a functional RNA splice site at the end of their coding regions. Our data show that a single functional J is located 1.3 or 1.6 kb upstream of each of the C lambda gene segments known to encode the Mcg, Kern- Oz-, and Kern- Oz+ isotypes. Therefore, the gene organization of this region of the human lambda locus is J1C1-J2C2-J3C3. The DNA sequences of J lambda 1, J lambda 2, and J lambda 3 presented in this paper establish that a single J lambda gene segment precedes each expressed C lambda gene segment, and support a model for the evolution of the human lambda JC clusters where J1C1 and J2C2-J3C3 arose from different ancestral JC units.

A single rearrangement event generates most of the chicken immunoglobulin light chain diversity

The chicken immunoglobulin lambda locus contains a single C lambda gene with a unique J lambda element, 1.9 kb upstream. The same V lambda gene (V lambda 1) is rearranged in most cells of the Bursa of Fabricius. This V lambda 1 gene is located, in germ-line configuration, 1.7 kb upstream from J lambda and in the same transcriptional orientation. Eight to twelve variable genes of the same set are found adjacent to the V lambda 1 gene, indicating that V-gene amplification did occur. Three of these genes were sequenced and proved to be pseudogenes, one of them having an inverted polarity. Data suggesting extensive somatic diversification of the V lambda 1 sequence are reported, including the possible use of nonfunctional V elements in a somatic gene-conversion-like process.

Variant (6 ; 15) translocation in a murine plasmacytoma occurs near an immunoglobulin kappa gene but far from the myc oncogene.

Chromosome translocations in B-lymphoid tumours are providing intriguing insights and puzzles regarding the role of immunoglobulin genes in the activation of the myc oncogene (reviewed in refs 1, 2). The 15 ; 12 translocations found in most murine plasmacytomas and the analogous 8 ; 14 translocation in human Burkitt's lymphomas involve scissions of murine chromosome 15 (human chromosome 8) near the 5' end of the c-myc gene and subsequent fusion near an immunoglobulin heavy-chain gene. The less well characterized 'variant' translocations found in about 15% of such tumours also involve the myc-bearing chromosome band, but exchange occurs with a chromosome bearing an immunoglobulin light-chain locus--in mice, the kappa-chain locus bearing chromosome 6 (refs 3-5) and, in man, chromosome 2 (or 22), at the same band at which the kappa (or lambda) locus lies (reviewed in ref. 1). The Burkitt variant translocations involve scissions 3' of c-myc; one 8 ; 22 translocation placed the C lambda locus just 3' of c-myc, but usually the chromosome 8 breakpoint is a greater, but unknown, distance away from c-myc, more than 20 kilobases (kb) in one 8 ; 2 translocation involving the C kappa gene. Little is known about the murine 6 ; 15 translocations, although a C kappa gene cloned from one plasmacytoma (PC7183) is linked, via chromosome 12 sequences, to an unidentified region of chromosome 15 (ref. 11). We describe here the chromosome fusion region from plasmacytoma ABPC4, which displays the typical reciprocal 6;15 translocations. We find that the chromosome 6 breakpoint is near C kappa but, unlike those in the heavy-chain locus, not at a position where immunoglobulin genes normally recombine. Moreover, the chromosome 15 sequences involved in the ABPC4 translocation are not derived from the vicinity of c-myc.

The isolation of a human Ig V lambda gene from a recombinant library of chromosome 22 and estimation of its copy number.

We report the first isolation and characterisation of a human Ig V lambda gene. The gene was isolated from a recombinant phage library of human chromosome 22 using a mouse Ig lambda cDNA as probe. DNA sequence analysis predicts a short leader peptide interrupted by an intron of 88 nucleotides, and a mature polypeptide of 96-97 amino acids which shares 61% homology with mouse V lambda I chains. Comparison with the amino acid sequence of known human lambda chains of all six subgroups shows agreement at 22/25 low variance positions. However the maximum homology with human chains is 49%, so we conclude that this sequence represents a new IgV lambda subgroup. The coding region is followed by the conserved heptamer, CACAGTG, and nonamer, ACATAAACC, sequences which have been implicated in V-J segment recombination. This gene has the hallmarks of an active V lambda gene including recently identified transcriptional controlling sequences. Probing genomic DNA with the subcloned V lambda gene detects a family of about 10 cross hybridizing members at low stringency and 2 at high stringency. There is limited polymorphism of the V lambda locus.

Rat c-myc oncogene is located on chromosome 7 and rearranges in immunocytomas with t(6:7) chromosomal translocation.

Two B-cell-derived tumours, human Burkitt's lymphoma (BL) and murine plasmacytoma (MPC), are regularly associated with a distinctive form of chromosomal translocation (for reviews see refs 1, 2). In BL, the distal portion of chromosome 8 breaks off and is transposed, in most cases, to chromosome 14, known to carry the immunoglobulin heavy-chain locus. In about 5% of the cases the same distal part of the chromosome 8 has moved to either chromosome 2 or 22, to the neighbourhood of the kappa or the lambda locus, respectively. In MPC the distal region of chromosome 15 is transposed to the chromosome 12, known to carry the immunoglobulin heavy-chain locus, or enters into reciprocal exchange with the kappa locus-carrying chromosome 6 (ref. 7). Several laboratories have located c-myc, the cellular homologue of the MC29 retroviral oncogene v-myc, to human chromosome 8 (refs 8-10) and mouse chromosome 15 (refs 11-13). It has also been shown that the BL- and MPC-associated translocations remove the c-myc gene from its original site and transpose it into or close to one of the immunoglobulin gene clusters. In view of the above findings we also looked for possible involvement of the c-myc gene in a B-cell-derived tumour of a third species, the rat. Rat immunocytomas of spontaneous origin carry a reciprocal translocation between chromosomes 6 and 7 (ref. 17). Here we have localized the c-myc locus to chromosome 7 of the rat. Moreover, we have found that the c-myc gene was rearranged in four of five immunocytomas carrying the characteristic chromosomal translocation.

Variable amplification of immunoglobulin lambda light-chain genes in human populations.

The human lambda immunoglobulin locus displays a series of restriction fragment length polymorphisms that are readily detected in small populations of normal individuals. Similar polymorphisms appear in populations of wild mice, suggesting that the lambda locus is subject to rapid variation within a single species. Here we show that the polymorphisms seen in the human lambda locus seem to have arisen from unequal meiotic crossing over, altering the number of lambda from as few as six to as many as nine per haploid genome. This expansion and contraction in the number of human lambda genes is significant in that it may affect an individual's capacity to produce variation among lambda light chain genes.

Amplification of immunoglobulin lambda constant genes in populations of wild mice.

The lambda immunoglobulin light chain (Ig lambda) locus of BALB/c inbred mice consists of two variable region gene segments (V lambda)1-3, and four constant region gene segments (C lambda)1,2,4,5. Each C lambda gene segment is associated with a unique joining segment (J lambda)2,4-7, and they are organized in two paired units, J3C3-J1C1 and J2C2-J4C4 (refs 4, 8). Using cDNA probes specific for C lambda 1 and C lambda 2 (ref. 9) we have analysed the genomic organization of the C lambda gene segments in wild-derived and inbred strains of mice. Although Southern blots of the genomic DNA of inbred mice show a constant pattern of hybridization, wild-derived mice show a high degree of variation in the number, size and intensity of hybridizing fragments. We have now found that, per haploid genome, mice of a Mus musculus musculus stock isolated from Sladeckovce, Czechoslovakia (CzII) have at least 12 C lambda segments, and mice of a Mus musculus domesticus stock 'Centreville Lights' from Centreville, Maryland (CL) have at least 8 C lambda segments. There appears to have been relatively recent amplifications of the C lambda gene segments in wild mice.