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V L Karpov

Publications and source records attributed to V L Karpov.

At least 19 recordsLinked to original sources
PubMed2006

RNA-protein interactions at the initial and terminal stages of protein biosynthesis as investigated by Lev Kisselev (on the occasion of his 70th anniversary).

This review highlights studies by Lev L. Kisselev and his colleagues on the initial and terminal stages of protein biosynthesis, which cover the period of the last 45 years (1961-2006). They investigated spatial structure of tRNAs, structure and functions of aminoacyl-tRNA-synthetases of higher organisms, and the final step of protein synthesis, termination of translation. L. Kisselev and his team have made three major contributions to these fields of molecular biology; (i) they proposed the hypothesis on the role of anticodon triplet of tRNA in recognition by cognate aminoacyl-tRNA synthetase, which has been experimentally confirmed and is now included in textbooks; (ii) identified primary structures and functions of two eukaryotic protein factors (eRF1 and eRF3) playing a pivotal role in translation termination; (iii) characterized a structural basis for stop codon recognition by eRF1 within the ribosome and discovered the negative structural elements of eRF1, limiting its recognition of one or two stop-codons.

Amino Acyl-tRNA Synthetases↗
PubMed2000

Tramtrack protein-DNA interactions. A cross-linking study.

Interaction of the Tramtrack protein from Drosophila melanogaster with DNA was analyzed by a cross-linking method. Tramtrack residues cross-linkable to the partially depurinated DNA were identified by direct sequencing. The N-terminal alpha-amino group of the protein DNA-binding domain was found to be the major product of cross-linking. The location of the N terminus on the DNA was determined by identification of the DNA bases that were cross-linked to the protein alpha-amino group. We conclude that accessory N-terminal peptide preceding the first zinc finger of Tramtrack directly interacts with DNA, both in specific and nonspecific DNA-protein complexes. Our finding explains the role in the protein binding of the DNA bases outside of the direct interaction with the zinc fingers.

Amino Acid Sequence↗
PubMed1996

Mapping of DNA-binding proteins along the yeast genome by UV-induced DNA-protein crosslinking.

UV-induced crosslinking of DNA-binding proteins to DNA in intact nuclei of Saccharomyces cerevisiae and subsequent 'protein image' hybridization were applied to map non-histone proteins along single-copy genes of yeast. We detected two polypeptides that most probably correspond to core subunits of yeast RNA-polymerase II in the coding region of transketolase gene (TKL2). Several non-histone proteins were also detected which bind to the upstream region of TKL2 gene, and to the intergenic spacer between calmodulin (CMD1) and beta-mannosyl transferase (ALG1) genes.

Calmodulin↗
PubMed1996

Mapping protein-DNA interactions with CIS-DDP: chromatin structure of promoter region of D. Melanogaster hsp 70 gene.

DNA-protein crosslinking by cis-dichlorodiammineplatinum (II) (cis-DDP) was applied to study chromatin structure in situ. Histone H1 (H5) is crosslinked to DNA in significant amounts whereas core histones remain practically unattached. "Protein image hybridization" experiments show that the 5'-region of the D metanogaster hsp 70 gene is free of histone H1 in both control nuclei and nuclei isolated from heat-shocked embryos.

Animals↗
PubMed1993

Identification of DNA binding proteins in vaccinia virus by DNA-protein crosslinking.

DNA binding proteins of vaccinia virus (VV) virions, strain LIVP, have been studied by their covalent crosslinking to DNA, using two-dimensional gel retardation electrophoresis of crosslinked DNA and proteins as well as the 'protein image' hybridization assay. Five proteins with molecular masses of 16, 25, 27, 41 and 54 kDa, respectively, associated with all analysed DNA sequences, including early and late genes and their promoters, have been identified.

Cross-Linking Reagents↗
PubMed1993

Mapping and positioning DNA-binding proteins along genomic DNA. Structure of D. melanogaster ribosomal 'Alu-repeats' and 1.688 satellite chromatin.

Chromatin structure of so-called 'Alu-repeat' in D. melanogaster ribosomal non-transcribed spacer that contains sequences homologous to the promoter of ribosomal genes has been studied. Using the 'protein image' hybridization assay based on UV-light-induced DNA-protein crosslinking and 2-D gel retardation electrophoresis, two proteins of the molecular mass of 50 kD (rABP50) and 70 kD (rABP70), associated with 'Alu-repeat' DNA have been found. Exo III mapping of crosslinking sites and DNase I footprinting have provided a detailed map of H1, rABP50 and rABP70 contacts within the 'Alu-repeat' and H1 and a non-histone protein contacts on satellite DNA. These data indicate precise positioning of non-histone proteins, histone H1 and nucleosomes within genomic regions studied and account for the presence of unusual 240 bp long nucleosomal particles in 'Alu-repeats'. The same approach can be adapted for successive mapping and positioning proteins on genomic DNA.

Animals↗
PubMed1993

Two non-histone proteins are associated with the promoter region and histone HI with the transcribed region of active hsp-70 genes as revealed by UV-induced DNA-protein crosslinking in vivo.

We described here an approach for mapping proteins on any sequence of genomic DNA. UV-induced DNA-protein crosslinking within whole cells and the 'protein image' hybridization technique (1) were applied to test the proteins bound to different regions of the D. melanogaster hsp-70 gene. The histone H1-DNA association with the coding region is shown to be maintained, even during very intensive transcription, but is absent in the promoter. Two non-histone proteins with apparent molecular masses of 50 kD (p50) and 100 kD (p100) are crosslinked only to the active hsp-70 gene regulatory region and preferentially bind to its complementary and coding DNA strands, respectively.

Animals↗
PubMed1990

Chromatin structure of Drosophila melanogaster ribosomal genes.

The chromatin structure of ribosomal genes of D. melanogaster has been studied by crosslinking proteins to DNA. We found that a number of histone contacts with DNA through histidine in the approximately 1 kb-long region surrounding the transcription initiation site, coding regions and the region of 240 bp-long repeats from the intergenic spacers (Alu-repeats) were weakened as compared to the inactive chromatin of the type II insertion. A protein with the molecular mass of 50 kDa (p50), associated with all DNA sequences analysed, has been discovered. Another protein with molecular mass of about 70 kDa (p70) has been found to be specific only for the Alu-repeats.

Animals↗
PubMed1989

Change in the pattern of histone binding to DNA upon transcriptional activation.

Patterns of histone binding to DNA of transcriptionally active D. melanogaster hsp70 genes within the nuclei have been analyzed by two methods of histone-DNA chemical cross-linking. When cross-linking is restricted to the central, "globular" regions of histones, it drops most for H1, to an intermediate extent for H2A and H2B, and least for H3 and H4 in transcriptionally active versus transcriptionally silent chromatin. When it occurs via histone terminal regions as well, cross-linking is quantitatively similar for active and inactive chromatin. Neither cross-linking method detects histones on the hsp70 promoter region. It appears that chromatin activation decreases histone binding to DNA via the "globular" regions, known to be essential for the folding of nucleosomes and the 30 nm chromatin fibril, but does not significantly affect the interaction of flexible and loosely bound histone "tails" with DNA. The role of these histone-DNA interaction changes in the unfolding of active chromatin and RNA polymerase reading through histone-bound DNA is discussed.

Animals↗
PubMed1984

Chromatin structure of hsp 70 genes, activated by heat shock: selective removal of histones from the coding region and their absence from the 5' region.

The presence of histones in hsp 70 genes was studied by "protein-image" hybridization technique after crosslinking histones to DNA. With increasing transcription of the genes, the coding region was demonstrated to be depleted first of H1 and then of all histones. This probably accounts for unraveling the 25 nm silent chromatin fiber to moderately and actively transcribed 10 nm fiber and linearized DNA. No histones were found in the 5'-terminal DNAase I-hypersensitive region, which may be a prerequisite to gene activation. The absence of histones on DNA correlates well with the high nuclease sensitivity and disappearance of the regular pattern in micrococcal nuclease digests of chromatin.

Animals↗