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D A Maslov

Publications and source records attributed to D A Maslov.

50 records · Page 3Linked to original sources

[Restriction map of maxicircle kinetoplastic DNA of Leishmania gymnodactyli].

Kinetoplast DNA (kDNA) of Leishmania gymnodactyli, as in other Trypanosoma species, is a complicated associate consisting of mini- and maxicircular molecules. Minicircular DNA are presented by the homogeneous in size and heterogeneous in base sequence population. Their size determined by the agarose electrophoretic mobility in gel is 920 pairs of nucleotides. Maxicircular molecules of DNA are homogeneous and contains about 38 thou pairs of nucleotides. The restriction map of maxicircular kDNA molecule of L. gymnodactyli has been made up on the basis of the cleavage data of the whole associate kDNA by restrictases.

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Conservative and divergent base sequence regions in the maxicircle kinetoplast DNA of several trypanosomatid flagellates.

The structure of kinetoplast maxicircle DNA from trypanosomatids Crithidia oncopelti, C. luciliae, Leptomonas pessoai and Leishmania gymnodactyli was compared by the blot hybridization method. The sizes of these molecules are 24.5, 34, 31 and 38 kilobase pairs (kbp), respectively. Labelled maxicircle fragments from C. oncopelti were used as probes. A general model of the structural organization of maxicircles is proposed according to which this molecule is composed of a 17kbp conservative region and a divergent one occupying the rest of the molecule. The conservative region contains the sequences homologous to those in all trypanosomatids. The sequence of the divergent region exhibits no cross homology detectable by high stringency hybridization. The main size differences between the maxicircle molecules from different trypanosomatid species are explained by the length variability of their divergent regions.

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[Kinetoplast DNA of Crithidia oncopelti. Recognition of the principles of structural organization of minicircular DNA molecules].

The heterogeneity of the kinetoplast minicircular DNAs from Crithidia oncopelti was studied by electron microscopy and restriction analysis. The associate of kinetoplast DNA comprises five main classes of minicircles sizing 0.42, 0.51, 0.62, 0.78 and 0.83 microns. Examination of cleavage patterns revealed that each class of the minicircles is heterogeneous in base sequence, but there are restriction fragments of the same size common for all these classes. A model of structural arrangement of minicircles of Crithidia oncopelti was proposed. This model is based on the block structure of minicircular molecules. Each minicircle is composed of one variable block (V-block), 810 base pairs, and from 1 to 4 more conservative blocks (C-blocks) each containing 445 base pairs.

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[Polypeptide synthesis dependent on kinetoplast DNA from Crithidia oncopelti in a cell-free system of E. coli].

The ability of a complex associate made up of maxi- and mini-circular molecules of kinetoplast DNA from C oncopelti to perform polypeptide synthesis in a cell-free system of conjugate transcription and translation of E. coli was demonstrated. The coding function of maxi-circular molecules and genetic inertness of mini-circular molecules of kinetoplast DNA during polypeptide synthesis in vitro was found. Total RNA from the kinetoplasts was shown to undergo translation in cell-free protein-synthesizing system of E. coli to form 9-10 polypeptides. The molecular weights of many polypeptides, whose synthesis in this system is performed by DNA and RNA from Kinetoplasts were found to be similar.

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[Presence of genes of ribosomal and transfer RNAs in kinetoplast DNA from two Crithidia species].

The coding properties of kinetoplast DNA from two respresentatives of the order Kinetoplastidae--Crithidia oncopelti and C. fasciculata--were studied. Experiments on hybridization of the whole network and fraction of minicircles with labelled 23S and 16S rRNA and with tRNA isolated from kinetoplasts of C. oncopelti clearly demonstrated the presence of the genes for these RNAs in the whole network and their absence in the minicircles. It may be thus concluded that the genes of ribosomal and transfer RNAs are localized in the maxicircular molecules. Similar efficiency of hybridization of rRNAs from C. oncopelti with kDNA from C. fasciculata and C. oncopelti revealed significant conservativity of ribosomal genes in the protozoa under study.

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Monophyly of endosymbiont containing trypanosomatids: phylogeny versus taxonomy.

To obtain additional information on the phylogenetic relationships within the family Trypanosomatidae (order Kinetoplastida), we have sequenced the small subunit ribosomal RNA genes from the endosymbiont containing species Herpetomonas roitmani TCC080, Herpetomonas sp. TCC263, Crithidia oncopelti ATCC 12982 and a partial large subunit rRNA gene from H. roitmani. The small subunit sequences in the two isolates of Herpetomonas are very similar but not identical, and so are their restriction digest profiles of kinetoplast DNA. The size of minicircles in both isolates is 4.2 kilobases. The inferred ribosomal RNA phylogenetic trees shows the genera Herpetomonas and Crithidia as polyphyletic. Endosymbiont-bearing herpetomonads cluster with the endosymbiont-bearing crithidias and a blastocrithidia to form a monophyletic clade, whereas the endosymbiont-free members of these genera are found elsewhere in the tree. These data support the hypothesis of a monophyletic origin of endosymbiosis in trypanosomatid evolution and also suggest that a taxonomic revision is needed in order to better describe the natural affinities in this family.

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Diversity and phylogeny of insect trypanosomatids based on small subunit rRNA genes: polyphyly of Leptomonas and Blastocrithidia.

With the aim of further investigating phylogenetic relationships in insect trypanosomatids, we have determined the sequences of small subunit rRNA genes from ten isolates, which were originally classified as Leptomonas, Blastocrithidia, and Wallaceina based on their morphology in the hosts. The inferred maximum likelihood, parsimony, and distance trees indicate that the Leptomonas and Blastocrithidia are polyphyletic, and confirm the polyphyly of Herpetomonas and Crithidia. Blastocrithidia triatoma and Leptomonas collosoma were among the earliest branching lineages among the insect trypanosomatids, while most other isolates were found within a closely related terminal clade, which also included Crithidia fasciculata. This analysis has clearly demonstrated that the morphological classification system of insect trypanosomatids does not always reflect their genetic affinities warranting its revision in the future.

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Diplonema spp. possess spliced leader RNA genes similar to the Kinetoplastida.

The phylogenetic placement of the genus Diplonema in relation to fellow phylum members Euglena and Trypanosoma has been uncertain. The spliced leader RNA gene, present in the euglenids and kinetoplastids in distinct forms, was a potential target for resolving this question. The first indication supporting a closer relationship to the kinetoplastids was the recognition of potential spliced leader RNA exon sequences in the genomic DNA of two Diplonema isolates. Examination of total cell RNA revealed transcripts in the anticipated size range at approximately 120 and 130 nt. Specific PCR amplification of a spliced leader RNA gene repeat was performed. The hallmark features of the kinetoplastid-type spliced leader RNA, specifically the 39-nt exon, splice-donor site, Sm-binding site and poly-T tract and the potential to form the requisite stem-loop structures, were found. Diplonema spp. are different from the kinetoplastids by virtue of C residues at positions 4 and 18 in the exon. While the intergenic spacer regions varied in size, each contained the complete sequence or remnants of a 5S ribosomal RNA gene. Possession of a functional spliced leader RNA gene of the kinetoplastid variety in Diplonema supports a closer evolutionary relationship with this group than with the euglenids.

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[Analysis of the sequence of repeats in divergent regions of maxi-circular DNA from kinetoplasts of Crithidia oncopelti].

The nucleotide sequence of direct clustered repeats from the divergent region of the maxicircle kinetoplast (mitochondrial) DNA from the protozoan Crithidia oncopelti was analysed. 10 kbp long divergent region contains 3 clusters composed of 18-23 tandem repeats of 82-83 bp (Sau-repeats) and a single cluster of five 417 bp repeats (EcoRI-repeats). The clusters are interspersed between the regions of nonrepetitive DNA. The details of the structural organization of repeats and their clusters were considered on the nucleotide sequence level. The possible ways of origin of the clusters are discussed.

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[Various elements of the structure of mini-ring kinetoplast DNA of Crithidia fasciculata].

The nucleotide sequence of 1.4 kbp SmaI-fragment of minicircle DNA from kinetoplasts of Crithidia fasciculata has been determined and some sequence elements characterized. The sequence contains several oligo(dT)blocks located on the same strand in phase with a period of DNA helix turn, thus representing a "bent helix". Both sides of the bent helix region are flanked by sequences capable of forming a cloverleaf structure. There are also two direct 150 bp repeats located 180 degrees apart on the circular map of the molecule. Each repeat contains the sites of H-strand and L-strand replication origin. The specific stem-loop secondary structure may be folded by the nucleotide sequence within the origins region. The alignment of the sequence determined with two other C. fasciculata minicircle sequences spanning over the bent helix and the adjacent regions has indicated the presence of several conserved sequence blocks, one of them representing the sequence of the bend. The divergence of three sequences occurred mainly by small insertions-deletions. Several open reading frames were found, the largest of which being capable of coding for the approximately 200 amino acids polypeptide.

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[Physicochemical properties of temperate phage DNA and its possible effect on the variability of Mycobacterium lacticolum].

The temperate phage 104 S was isolated from the S variant of Mycobacterium lacticolum, strain 104, and some of its characteristics were studied. The content of GC pairs in the phage DNA was 77 mole% as was calculated from the melting profile or 65 mole% as was calculated from the value of buoyant density in CsCl. The DNA was shown to be composed of 18,000 nucleotide pairs. DNA restriction fragments of M. lacticolum R, S and M variants were subjected for the first time to molecular hybridization with [32P]DNA of the temperate phage. The genome of the three M. lacticolum variants and the genome of a non-dissociating S variant clone were shown to contain sequences homologous to the DNA sequence of phage 104 S. Differences are found among the variants in the hybridizing DNA fragments. These data indicate that the phage DNA may actively be involved in the variability of the culture. Its participation can be realized by the different mode of prophage incorporation into the genome of the variants.

Base Composition↗

[Characteristics of kinetoplastic DNA from Leptomonas pessoai].

The kinetoplast (mitochondrial) DNA from trypanosomatid Leptomonas pessoai represents a network, composed of mini-circles heterogeneous in base sequence and homogeneous maxi-circles and thus has the main structural features in common with DNAs from kinetoplasts of other Trypanosomatidae. The size of mini-circular molecules of DNA is 1,35 kilobase pairs (kbp) and that of maxicircular molecules-30,9 kbp. Based on the data of single and double restriction cleavages the physical map of the maxi-circular molecules was constructed for the endonucleases BamHI, BglII, BspI , HindIII, MspI, SalGI and PstI.

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