Somatic hypermutagenesis in immunoglobulin genes. II. Influence of neighbouring base sequences on mutagenesis.

A new approach for the analysis of hotspots of mutations is described. It is based on the classification of hotspot site sequences. Using this approach, the consensuses RGYW and TAA of hotspot sites were revealed in the V gene. Correlation between somatic mutations and these consensuses is investigated by the statistical weight method in 323 somatic substitutions in 14 V genes. Assuming the absence of any correlation, the probability of observing such data in the sample would be very low (0.0003). These results support the idea that emergence of somatic mutation is significantly influenced by neighbouring base sequences. This idea was also supported by the analysis of 296 somatic mutations in flanking sequences of V genes. It is supposed that this influence is an important feature of somatic hypermutagenesis.     

Characterization of several LINE-1 elements in Microtus kirgisorum

Several LINE-1s have been isolated and characterized from genomic DNA of the vole, Microtus kirgisorum. Blot hybridization revealed specific restriction patterns of L1 elements in vole genomes. Rehybridization of the genomic blot with a cloned 5′-end fragment revealed two major bands indicating the presence of two different L1 subfamilies. The copy numbers are estimated for different parts of M. kirgisorum L1 elements. Data also demonstrate that most vole L1 elements are truncated at the 5′-end; however, in contrast to mouse, the ORF1 copy number is higher in vole. A difference between the substitution rates of the ORF1 5′-region (approximately 330 nucleotides) and the rest of the L1 coding regions is revealed. Received: 12 January, 1999 / Accepted: 18 March, 1999     

Regression trees for analysis of mutational spectra in nucleotide sequences.

MOTIVATION: The study and comparison of mutational spectra is an important problem in molecular biology, because these spectra often reveal important features of the action of various mutagens and the functioning of repair/replication enzymes. As is known, mutability varies significantly along nucleotide sequences: mutations often concentrate at certain positions in a sequence, otherwise termed 'hotspots'. RESULTS: Herein, we propose a regression analysis method based on the use of regression trees in order to analyse the influence of nucleotide context on the occurrence of such hotspots. The REGRT program developed has been tested on simulated and real mutational spectra. For the G:C-->T:A mutational spectra induced by Sn1 alkylating agents (nine spectra), the prediction accuracy was 0. 99. AVAILABILITY: The REGRT program is available upon request from V.Berikov.     

Effect of winter conditions on distributions of anoxic phototrophic bacteria in two meromictic lakes in Siberia,Russia

The year-to-year variations of vertical distribution and biomass of anoxic phototrophic bacteria were studied during ice periods 2003–2005 and 2007–2008 in meromictic lakes Shira and Shunet (Southern Siberia, Russian Federation). The bacterial layers in chemocline of both lakes were sampled with a thin-layer hydraulic multi-syringe sampler. In winter, biomass of purple sulphur bacteria varied considerably depending on the amount of light penetrating into the chemocline through the ice and snow cover. In relatively weakly stratified, brackish Shira Lake, the depth of chemocline varied between winters, so that light intensity for purple sulphur bacteria inhabiting this zone differed. In Shira Lake, increased transparency of mixolimnion in winter, high chemocline position and absence of snow resulted in light intensity and biomass of purple sulphur bacteria exceeding the summer values in the chemocline of the lake. We could monitor snow cover at the lake surface using remote sensing and therefore estimate dynamics and amount of light under ice and its availability for phototrophic organisms. In Shunet Lake, the light intensities in the chemocline and biomasses of purple sulphur bacteria were always lower in winter than in summer, but the biomasses of green sulphur bacteria were similar.     

Organization and chromosomal localization of a B1-like containing repeat of Microtus subarvalis

A repetitive DNA sequence, MS2, was isolated from EcoRI-digested genomic DNA of the vole, Microtus subarvalis. The fragment was cloned and sequenced. Sequence analysis of this 1194-bp fragment revealed a 156-bp region demonstrating a 55% homology with the mouse B1 repeat. The remaining MS2 sequence shows no significant homology with other known GenBank sequences. The results of in situ hybridization of MS2 on vole metaphase chromosomes indicate the fragment is confined to heterochromatin blocks of the sex chromosomes in all but one species (M. arvalis). Distribution of MS2 sequences provides evidence for heterogeneity of the giant heterochromatin blocks of the XY Chromosomes (Chrs) in voles, for the unique cluster-like localization of MS2 within these blocks. Received: 10 October 1995 / Accepted: 30 March 1996     

GeneBuilder: interactive in silico prediction of gene structure.

MOTIVATION: Prediction of gene structure in newly sequenced DNA becomes very important in large genome sequencing projects. This problem is complicated due to the exon-intron structure of eukaryotic genes and because gene expression is regulated by many different short nucleotide domains. In order to be able to analyse the full gene structure in different organisms, it is necessary to combine information about potential functional signals (promoter region, splice sites, start and stop codons, 3' untranslated region) together with the statistical properties of coding sequences (coding potential), information about homologous proteins, ESTs and repeated elements. RESULTS: We have developed the GeneBuilder system which is based on prediction of functional signals and coding regions by different approaches in combination with similarity searches in proteins and EST databases. The potential gene structure models are obtained by using a dynamic programming method. The program permits the use of several parameters for gene structure prediction and refinement. During gene model construction, selecting different exon homology levels with a protein sequence selected from a list of homologous proteins can improve the accuracy of the gene structure prediction. In the case of low homology, GeneBuilder is still able to predict the gene structure. The GeneBuilder system has been tested by using the standard set (Burset and Guigo, Genomics, 34, 353-367, 1996) and the performances are: 0.89 sensitivity and 0.91 specificity at the nucleotide level. The total correlation coefficient is 0.88. AVAILABILITY: The GeneBuilder system is implemented as a part of the WebGene a the URL: http://www.itba.mi. cnr.it/webgene and TRADAT (TRAncription Database and Analysis Tools) launcher URL: http://www.itba.mi.cnr.it/tradat.     

Microbial community of the chemocline of the meromictic Lake Shunet (Khakassia,Russia) during summer stratification

The spatio-temporal organization of the bacterial community inhabiting the chemocline of the stratified meromictic Lake Shunet (Khakassia, Russia) was investigated from May to September 2005 by means of microscopy, analysis of photosynthetic pigments, and PCR-DGGE with subsequent 16S rDNA analysis. The samples were collected with a multisyringe stratification sampler, sampling being performed every 5 cm. It was demonstrated that, during the period of investigation, there were no large changes in the bacterial community of the chlemocline, at least among the detected forms. During the whole period of study, purple sulfur bacteria related to Lamprocystis purpurea (Chromatiaceae) were predominant in the chemocline. Beneath the layer of purple bacteria, green sulfur bacteria were revealed that were phylogenetically distant from strain ShNPel02, which was previously isolated from this lake. Development of phytoflagellates of the genus Cryptomonas was observed in the upper zone of the chemocline. In the chemocline of Lake Shunet, the numbers of picoplankton cyanobacteria of the genus Synechococcus increased from May to September. It was demonstrated that the application of universal bacterial primers for DGGE resulted in the same qualitative distributional pattern of predominant species as microscopic studies.