Stage transitions in B-lymphocyte differentiation correlate with limited variations in nuclear proteins.

Rabilloud, Thierry; Pennetier, Jean Louis; Hibner, Urszula; Vincens, Pierre; Tarroux, Philippe; Rougeon, François

doi:10.1073/pnas.88.5.1830

Cited by 16 publications

(13 citation statements)

References 34 publications

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“…These conclusions are in agreement with the results showing that a very limited number of nuclear proteins mark the stages competent for immunoglobulin gene rearrangement in B-cell development (35).…”

supporting

confidence: 82%

Three lymphoid-specific factors account for all junctional diversity characteristic of somatic assembly of T-cell receptor and immunoglobulin genes.

Kallenbach

Doyen

d'Andon

et al. 1992

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

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The somatic diversity immunglobulin and T-cell receptor diversity is largely provided by the junctional variation created during site-specific rearrangement of separately encoded gene segments. Using a transient transfection assay, we demonstrate that the recombination activating genes Ragl and Rag2 direct site-specific rearrangement on an artificial substrate in poorly differentiated as well as in differentiated nonlymphoid cell lines. In addition to a high frequency of precise recombination events, coding joints show deletions and more rarely P-nucleotide insertions, reminiscent of immunoglobulin and T-cell receptor junctions found in fetal tissues. N-region insertions, which are characteristic of adult junctional diversity, are obtained at high frequency upon transfection of a terminal deoxynucleotidyltransferase expression vector together with Ragl and Rag2. These results show that only three lymphoid-specific factors are needed to generate all types ofjunctional diversity observed during lymphoid development.Immunoglobulin (Ig) and T-cell receptor (TCR) gene assembly is achieved through site-specific recombination events, from separately encoded variable (V), in some cases diversity (D), and junction (J) gene segments. Much of the immunoglobulin and TCR diversity is generated by the combinatorial rearrangement of a large number of V, D, and J gene segments (for a review see refs. 1 and 2).Recombination signal sequences (RSSs) situated adjacent to each gene segment provide the targets for recombination. RSSs are composed of a palindromic heptamer and an (A+T)-rich nonamer separated by a spacer of 12 or 23 base pairs (bp) (3). Rearrangement only occurs between RSSs with spacers of different length. RSSs are sufficient to target rearrangement of artificial substrates containing no other antigen receptor sequences (4-7).Two main types of joints are formed during the recombination process: coding joints created by the juxtaposition of the gene segments and reciprocal joints by contiguous RSSs (8). Whereas the heptamers in the reciprocal joints are generallyjoined back to back without nucleotide insertions or deletions, the coding joints are subjected to extensive processing (8). The junctions formed during rearrangement constitute another source of diversity. Several nucleotides can be deleted and two types of insertions can be found. Random nucleotide additions, resulting in N-region insertions, are thought to be introduced by terminal deoxynucleotidyltransferase (TdT) (9-11). P-nucleotide insertions represent the inverted repeat of the adjacent coding sequence. Their addition has been proposed to be a compulsory step of the recombination mechanism (12).Genomic DNA transfection experiments have allowed the isolation of two recombination activating genes, Ragl and Rag2 (13-15). Together these genes are able to confer sitespecific recombination activity to NIH 3T3 fibroblasts. Although it is not formally excluded that they activate other genes that would be responsible for the recombination activity, Ragl...

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supporting

confidence: 82%

Three lymphoid-specific factors account for all junctional diversity characteristic of somatic assembly of T-cell receptor and immunoglobulin genes.

Kallenbach

Doyen

d'Andon

et al. 1992

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

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“…Principal component analysis was performed on the 24 autoradiograms to evaluate the relatedness of the 24 gels based on the relative abundance of each protein (Tarroux 1983;Tarroux et al 1987;Lefkovits et al 1988;Rabilloud et al 1991). This mathematical analysis allows the representation of objects defined by a set of variables.…”

Section: Methodsmentioning

confidence: 99%

“…the intensity of labeling). This method has proved to be a good tool for defining the relatedness of a set of two-dimensional gels Lefkovits et al 1988;Rabilloud et al 1991). The relations between the 24 autoradiograms were represented in axes 2 and 4 which contained respectively 9.4% and 4.5% of the information.…”

Section: Global Analysis Of Two-dimensional Gels Leaf Proteins Ofmentioning

confidence: 99%

Characterization of chilling-acclimation-related proteins in soybean and identification of one as a member of the heat shock protein (HSP 70) family

Cabané

Calvet

Vincens

et al. 1993

Planta

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Through a 5-d exposure at 14 degrees C/8 degrees C (day/night), soybean (Glycine max [L.] Merr.) was acclimated to a lower temperature of 8 degrees C. In order to assess changes in protein synthesis related to chilling acclimation, proteins were labeled in vivo with [35S]methionine, separated by two-dimensional gel electrophoresis, and the derived autoradiograms were subjected to computer analysis. Two sets of chilling-acclimation-related proteins were characterized following exposure and labeling at 8 degrees C. One set corresponded to proteins whose synthesis was stimulated in acclimated plants in comparison with non-acclimated plants after transfer to 8 degrees C for 2 d. The other set also displayed an enhanced synthesis in the acclimated plants versus the non-acclimated plants but after 7 d of exposure at 8 degrees C. Most of these chilling-acclimation-related proteins were not increased during the acclimation period at 14 degrees C. Using microsequence analysis, one of these proteins was shown to have a high sequence homology with members of the heat-shock protein (HSP 70) family.

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“…In case of different inheritance, the heterozygote can be found on any point of the hypervolume (a mathematical 'simplex') where the 2 vertices correspond to all possible combinations of complete dominance or recessivity for the n proteins. In the context of combinatorial or synergistic gene regulation, where the specificity of expression arises from the coordinate presence of two or more factors of restricted specificity (Yamamoto, 1985;Rabilloud et a!. 1991), this geometrical description may be useful to realize the extent to which an heterozygote, compared to its parents, can display unpredictable patterns of genetic expression whenever different types of inheritance result in a particular distribution of factors.…”

Section: Discussionmentioning

confidence: 99%

Quantification of dominance for proteins pleiotropically affected by opaque-2 in maize

Damerval

Vienne

1993

Heredity

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The maize opaque-2 (o2) mutation is known to have numerous pleiotropic effects. Using two different genetic backgrounds, endosperm proteins from isogenic lines of the o2 locus and their reciprocal hybrids were compared by high-resolution two-dimensional electrophoresis (2-D PAGE), and the 2-D gels stained with Coomassie blue or silver were analysed using a computerassisted system for quantification of polypeptide spots. The aim of this work was (i) to identify polypeptides, the amount of which is affected by the single gene substitution, and (ii) to assess the dominance relations for every affected polypeptide. The triploid nature of the endosperm allowed the effects of 0, 1, 2 and3 doses of the wild-type 02 allele to be examined. When using a basic pH range in the first dimension (IEF), we observed the known effects of o2 on prolamin storage proteins, mostly being 22 kDa, zeins. In a pH 5-7 IEF range, we detected 14 polypeptides whose amount is affected by the o2 mutation in both backgrounds. Eight of them were more abundant in the wild type than in the mutant and six were more abundant in the mutant than in the wild type. Dominance of the wild type allele was found for the amounts of 10 polypeptides, while the amounts of two of them displayed additive inheritance. This is an example where pleiotropically related characters display different inheritance, which could generate heterozygous phenotypes with original properties as compared to their homozygous parents. Because 02 belongs to the basic region-leucine zipper family of transcription activators, our results show that 2-D PAGE of isogenic lines represents a straightforward method to identify gene products affected by a transcription factor, whether they are products of its target genes, or products indirectly affected by far-reaching effects. This is a prerequisite to understanding the multiple molecular and physiological consequences of the action of such factors.Keywords: dominance, isogenic lines, maize, opaque-2, plelotropy. IntroductionNumerous single mutations are known that have multiple phenotypic consequences (Caspari, 1952;Wright, 1968). Various mechanisms can be put forward to explain pleiotropy. For example, a mutation of a gene coding for a trans-acting transcription factor binding to the promoter regions of different genes could result in variation of the characters they control (Hope & Struhl, 1985;Sassone-Corsi, 1985). At the post-translational level, it has been shown that differences in the degree of sialylation of multiple hydrolases could be due to the mutation of the gene coding for neuraminidase (Womack et at., 1981). Keightley & Kacser (1987) investigated a branched metabolic pathway where the two outputs represent two characters whose variation is affected by changes in any of the genetically specified enzymes in the system. However, characterization of the mutation may not be sufficient 38 to predict or understand all the diverse pleiotropic effects. Recent cloning of the r (rugosus) gene in pea revealed that it encodes a starch-br...

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Stage transitions in B-lymphocyte differentiation correlate with limited variations in nuclear proteins.

Cited by 16 publications

References 34 publications

Three lymphoid-specific factors account for all junctional diversity characteristic of somatic assembly of T-cell receptor and immunoglobulin genes.

Three lymphoid-specific factors account for all junctional diversity characteristic of somatic assembly of T-cell receptor and immunoglobulin genes.

Characterization of chilling-acclimation-related proteins in soybean and identification of one as a member of the heat shock protein (HSP 70) family

Quantification of dominance for proteins pleiotropically affected by opaque-2 in maize

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