Structural differences in a single gene encoding the V kappa Ser group of light chains explain the existence of two mouse light-chain genetic markers.

Abstract

Two phenotypic markers of mouse immuno- 2 for review). In several instances, the expression of strainspecific idiotypes has been shown to be due to differences among strains in the structure or repertoire of particular VH gene segments (3-5). A number of immunoglobulin K L-chain polymorphisms have been described (6-12), and in several instances Southern hybridization analyses have provided evidence that mice exhibit restriction enzyme polymorphisms involving VK gene segments (13-15).The present study explores the molecular genetic basis for two mouse VK polymorphisms, the IB-peptide marker (16) and the Efl isoelectric focusing marker (17). Both have been shown to be manifestations of the production of a characteristic group of K-chain V regions (called VKSer) by several strains of mice (C58/J, AKR/J, PL/J, and RF/J) but not by BALB/c and most other strains (18). Expression of these and other K polymorphisms maps to the K locus, which is closely linked to the Lyt-2 and Lyt-3 genes on chromosome 6 (19). Gottlieb et al. (18) have identified myeloma tumors producing K chains corresponding to the IB-peptide marker and Efl1 polymorphism in the C.C58 and C.AKR strains, which differ from BALB/c only in the region of the K and Lyt-2/Lyt-3 loci (20). These myelomas permitted isolation of a cDNA probe specific for the VKSer group, and Southern hybridizations with this probe provided evidence for a single strongly hybridizing VKSer-related gene in liver DNA of all strains tested (13). Restriction enzyme polymorphisms were observed among the strains tested, which correlated completely with expression or nonexpression of the IB-peptide and Efl1 phenotypic markers. Expressor strains contained the IgkVSera allele and nonexpressor strains contained the IgkVSerb allele (13,21).In the present studies, the VKSer-related genes of the expressor strain, C.C58, and the nonexpressor strain, BALB/c, have been cloned from phage libraries of liver DNA. Comparison of their nucleotide sequences demonstrates differences in their coding and 5' flanking regions, which explain the differences observed in expression of the IB-peptide and Efla phenotypic markers. Findings also bear generally upon the nature and origins of polymorphisms in mouse VK genes. MATERIALS AND METHODSCloning of C.C58 and BALB/c V,,Ser Genes. Liver DNA from C.C58 (20) and BALB/cAn mice raised in our colony was extracted according to a modification of the method of Maniatis et al. (22). DNA was subjected to partial digestion with Mbo I (New England Biolabs), and fragments 10-20 kilobases long were cloned into the BamHI site of the X phage vector EMBL3 (Promega Biotec, Madison, WI) according to Abbreviations: V and J, variable and joining regions of immunoglobulin; H chain and L chain, heavy chain and light chain. 9134The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

Conservation of sequence in recombination signal sequence spacers

“…Sequences were aligned using 'pileup', a GCG program that aligns groups of sequences based on comparison of the closest related pairs, and introduces gaps to promote optimal alignment. As experiments suggest that RSS function efficiently only if the first three nucleotides of the heptamer are fixed at CAC, and the heptamer and nonamer are separated by a spacer with variation in length of 11-13 bp or [22][23][24] bp only [7], gaps were inserted for optimal alignment based on these criteria. This was achieved using the pileup parameters 'gap weight' set at three, and 'gap length weight' set at 0.2.…”

Establishment and Functional Implications of B‐cell Connectivity

“…1 (indicated by boxes) compared to their most homologous sequences found in the literature. The 1524 is compared to the L7 germline sequence (Pech et al 1981), the 3339 to the 1210.7 sequence , the 4457 to the VKser germline sequence (Boyd et al 1986) and the 4340 to the PC 2880 sequence (Weigert et al 1978). All other designations are as in Fig.…”

MouseV k gene classification by nucleic acid sequence similarity

“…*References: 1, (Heinrich et al 1984); 2, (Clarke et al 1985); 3, (Shlomchik et al 1987c); 4, (Strohal et al 1987); 5, (Pech et al 1981); 6, (Altenburger et al 1980);7, (Eilat et al 1988); 8, (Kofler et al 1989); 9, (Sablitzky and Rajewsky 1984); 10, (Buckel et al 1987);11, (Even et al 1985);12, (H6chtl et al 1982); 13, (Kofler et al 1987b);14, (Kofler et al 1988); 15, ; 16, ; 17, (Shlomchik et al 1987a); 18, (Nahmias et al 1988);19, (Liu et al 1987);20, (Parslow et al 1984); 21, (Liu 1987); 22, (Kaartinen et al 1983); 23, (Griffiths et al 1984); 24, (Seidman et al 1978); 25, (Kelley et al 1985); 26, (Seidman et al 1979); 27, (Max et al 1980); 28, (Darsley and Rees 1985); 29, (Near and Haber 1989); 30, (Reininger et al 1987); 31, ; 32, (Campbell 1987); 33, (Meek et al 1989); 34, (Selsing and Storb 1981); 35, (Joho et al 1984); 36, (Lutz and Davie 1988); 37, (Corbet et al 1987); 38, (Borden and Kabat 1987);39, (Schiffet al 1983); 40, (Oilier et al 1985); 41, (Riley et al 1986); 42, (Kwan et al 1981); 43, (Boyd et al 1986); 44, …”

“…RFLP data suggested four to six ~19/28 germline genes (Kofler et al 1989), one of which, a F~28 germline gene, also known as VkSer, from haplotypes Igk-VSer ~, lgk-VSer b, lgk-VSe/, and Igk-VSer d, has been cloned (Boyd et al 1986, Ponath et al 1989. F~19/28 genes encoded antibodies to trinitrophenyl (Hawley et al 1982), carcinoembryonic antigen (Cabilly et al 1984, Beidler et al 1988, human breast/lung/colon cancer cells (Sahagan 1986), influenza hemagglutinin (Meek et al 1989), and an RNA-specific (Eilat et al 1988) and some RF-like autoantibodies (Kofler et al 1989, Shlomchik et al 1987a, 1987c.…”