ATP synthase from bovine mitochondria: sequences of imported precursors of oligomycin sensitivity conferral protein, factor 6, and adenosinetriphosphatase inhibitor protein

Oligomycin sensitivity conferral protein (OSCP), factor 6 (F6), and ATPase inhibitor protein are all components of the ATP synthase complex of bovine mitochondria. They are encoded in nuclear DNA. Complementary DNA clones encoding the precursors of these proteins have been isolated from a bovine library by using mixtures of synthetic oligonucleotides as hybridization probes, and their DNA sequences have been determined. The deduced protein sequences show that the OSCP, F6, and inhibitor proteins have N-terminal presequences of 23, 32, and 25 amino acids, respectively. These presequences are not present in the mature proteins. It is assumed that they serve to direct the proteins into the mitochondrial matrix. The cDNA clones have also been employed as hybridization probes to investigate the genetic complexity of the three proteins in cows and humans. These experiments indicate that the bovine and human inhibitor and bovine F6 proteins are encoded by single genes but suggest the possibility of the presence in both species of more than one gene (or pseudogenes) for the OSCP.     

Bacterial Communities of Regressive Spots in Ombrotrophic Bogs: Structure and Functions

Microbiology - This research analyses the structure and functions of bacterial communities of regressive spots in ombrotrophic bogs. Algal biomass was found to predominate in the biomass structure...     

Antitumor efficacy of combined gene and radiotherapy in animals

Antitumor efficacy of the combined suicide gene therapy and radiotherapy was studied on the model of CT26 murine colon adenocarcinoma. CMV-FCU1-IRES-mGM-CSF-pGL3 construct with PEG-PEI-TAT (FCU1–mGM/5-FC) block copolymer as a vector was used for intratumoral administration. Tumors were irradiated with a single 5 Gy dose. The efficacy was evaluated according to the grade of tumor growth inhibition (T/C) and lifespan of the animals. Pronounced antitumor activity of the combined use of FCU1–mGM/5-FC system with radiotherapy on the background of prolonged lifespan and the synergism of the applied methods was revealed.     

Functional diversity of ant-associated bacterial communities

It was found that ants significantly affect the physiological activity and functional diversity of soil microbial communities, and redistribution of biophilic elements (C and N) down through the profile occurs in anthills compared to the control soil, as well as their accumulation in the underground part of the ant nests. A high urease activity was revealed in ant nests and ants. Functional dissimilarities of bacterial communities in all studied objects were determined by the multisubstrate test.     

Oligonucleotide microarray for subtyping of influenza virus A neuraminidase

Microarray for influenza A neuraminidase subtyping was presented. Selection of oligoprobes proceeded in two steps. First step included selection of peptides specific for each subtype of neuraminidase. At the second step oligoprobes were calculated using found peptides structures with the subsequent additional selection of the most specific and representative probes. From 19 to 24 probes were used for determination of each subtype of neuraminidase. Microchip testing for 19 samples with the most widespread types (N1 and N2) specifies in unequivocal definition 18 of them and only one isolate has not been identified.     

Oligonucleotide conjugates with minor groove ligands as probes for hybridization microarray chips

A possibility of using oligonucleotide conjugates with minor groove ligands as probes for hybridization microarray chips was studied. The oligonucleotide conjugates contain a hairpin ligand (MGB) composed of two tripyrrolcarboxamide residues with an aminocaproic acid residue as a linker and bound to the oligonucleotide duplex AT tract in a site-specific manner. We used as (5'-3') probes GACAAGAp, GACAAAAp, GACAAGA-MGB, and GACAAAA-MGB. The oligonucleotides labeled with Cy3 cyanine dye, Cy3-ACTAATTTTGTC and Cy3-ACTAATCTTGTC, were used as targets. The maximal MGB effect on the fluorescence level of microarray chip spots, which caused its fourfold increase as compared with the initial unmodified duplex, was observed for the duplex containing only AT pairs in the ligand binding site. The presence of A-C and G-T mutations in the binding site (imperfect duplexes) or a C-G pair (perfect duplex) affects the change in fluorescence level to a considerably lesser degree.     

Study of genetic diversity in potato cultivars using PCR analysis of organelle DNA

The genetic diversity of 98 potato cultivars of Russian and foreign breeding was studied using of PCR with organelle-specific primers. The polymorphism of both plastid (atpE, trnG/trnK) and mitochondrial (rps, atp6) loci was revealed. Eight different haplotypes were detected in the sample of cultivars studied. Comparatively low of polymorphism of organelle DNA in the potato cultivars was demonstrated: most cultivars (91 or 92.9%) possessed only two haplotypes (I and II); 62 cultivars of them had similar "cultural" cytoplasmic type (haplotype I). The breeding cultivars of the Russian and foreign origin did not differ from each other in frequency of basic haplotypes.     

Tissue Specificity of Alternative Splicing of Transcripts Encoding Hampin,a New Mouse Protein Homologous to the <Emphasis Type="Italic">Drosophila</Emphasis> MSL-1 Protein

A number of mammalian genomes have one gene copy encoding the protein that we named hampin. A search in a number of databases revealed a distant homologue, the well-known Drosophila protein MSL-1 (male-specific lethal 1). An alternative splicing of mRNA led to a significant diversity of structural hampin variants with different domain compositions. We analyzed the tissue-specific expression of five mouse hampin variants using RT-PCR. Two variants encoding hampin proteins with truncated N termini were shown to have a restricted tissue specificity: they are exclusively expressed in the testes. The mRNAs of other hampin variants were detected in all the tested tissues at comparable levels. We obtained polyclonal antibodies to the recombinant hampin and used them to demonstrate that at least one of the variants is predominantly localized in the nucleus. The specific features of the hampin primary structure and its possible functions as a member of the hampin/MSL-1 family of proteins are discussed.__________Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 4, 2005, pp. 363–371.Original Russian Text Copyright © 2005 by Dmitriev, Pestov, Korneenko, Gerasimova, Zhao, Modyanov, Kostina, Shakhparonov.The article was translated by the authors.