[Isolation of total RNA from baker's yeast].

A simple method of production of total RNA from baker's yeast was developed. Total RNA was isolated from yeast (Saccharomyces cerevisiae) biomass using lysis with sodium dodecyl sulfate at 100 degrees C for 40-60 min and subsequent precipitation of the target product with 3 M NaCl. The preparation obtained was characterized in detail: yield of total RNA from 1 kg of pressed yeast, 9.

[Interaction of HIV-1 reverse transcriptase with new minor groove ligands and their conjugates with oligonucleotides].

The influence of new non-natural regular minor groove binders (MGB), containing 2-4 imidazole, pyrrole or thiazole residues, and their conjugates with oligonucleotides, on the polymerization reaction catalyzed by HIV-1 reverse transcriptase was analyzed. Various model template-primer complexes: poly(A)-oligo(U), poly(A)-oligo(dT), poly(dA)-oligo(U), poly(dA)-oligo(dT) and activated DNA were used. The concentration of oligopeptides, giving 50% inhibition (I50) of the RT-dependent polymerization reaction, was shown to depend strongly on the structure of template-primer complexes, number and type of the heterocycle rings in the MGBs analyzed.

[Interaction of proteins from general transcription complex RNA polymerase II with oligoribonucleotides].

We have analyzed an interaction of the general transcription complex RNA polymerase II proteins (RNA polymerase II, factors TBP, TFIIB, TFIIF, TFIIE and TFIIH) S. cerevisiae with the oligoribonucleotides. With the help of method EMSA was shown that labeled 32P labeled oligoribonucleotide 5'-ACUCUCUUCCGCAUCGC-3' (r-17) binds with the proteins and generates three species of the complexes with the three major shifts.

[The role of the 3'-CCA sequence in the interaction of tRNA(Phe) from E. coli and Thermus thermophilus with homologous phenylalanyl-tRNA-synthetases].

The 3'-CCA end of tRNA(Phe) from E. coli and Thermus thermophilus was modified by stepwise degradation and ligation of the shortened tRNA with different trinucleotides (pUpUpA, (pA)3, (pC)3, (pU)3). Kinetic parameters for the aminoacylation reaction of modified tRNAs have been determined.

[Structural elements of 80S ribosomes located near the 5'-region of the mRNA-binding center].

Affinity labeling of 80S ribosomes with 4-(N-2-chloroethyl-N-methylamino)benzylmethylphosphoramides of oligoribonucleotides [32P]AUGUn--mRNA analogs--was studied in three model complexes: 80S.ClRCH2N(CH3)-pAUGU6.Met(Phe)2-trRNA(Phe), 80S.

[Interaction of catalytically active antibodies with oligoribonucleotides].

The interaction of antibodies from blood sera of patients with autoimmune pathology, systemic lupus erythematosus with oligoribonucleotides was studied. The RNA-hydrolyzing activity was shown to be an intrinsic property of autoantibodies. Enzymic activity of antibodies in hydrolysis of poly(U) was estimated at 20-40% of that of RNase A.

[Affinity modification of 40S ribosomal subparticles from human placenta with mRNA analogs--AUGUnC oligoribonucleotide analogs with an alkylating group at the 3'-end].

Using 2',3'-O-[4-N-(2-chloroethyl)-N-methylamino]benzylidene derivatives of AUGUn[32P]pC (mRNA analogues), affinity labelling of human placenta 40S ribosomal subunits has been investigated in model initiation complexes obtained in the presence of the ternary complex eIF-2.GTP.Met-tRNA(fMet).

[Interaction of mRNA analogs--oligoribonucleotides AUGU3, AUGU6 and (pU)6 with 80S ribosomes from human placenta in the presence of related tRNA and a protein-synthesizing system].

Binding of oligoribonucleotides AUGUn (n-3 and 6) and (pU) to 80S ribosomes from human placenta in the presence of cognate tRNAs and a "ribosome-free" protein-synthesizing system from rabbit reticulocytes has been studied. The binding of the mRNA analogues resulted in formation of stable post-translocational complexes (which may be easily isolated by centrifugation in sucrose density gradient): 80S.AUGU3.

[Affinity modification of 40S ribosomal subparticles from human placenta by derivatives of oligoribonucleotides, containing an AUG codon. Ribosomal proteins, forming a codon-anticodon interaction region].

Using 2',3'-O-[4-N-(2-chloroethyl)-N-methylamino]benzylydene derivatives of AUGUn [32P]pC and 4-[N-(2-chloroethyl)-N-methylamino]benzylmethylphosphoamide derivatives of [32P]pAUGUn, the affinity labelling of human placental 40S ribosomal subunits was studied within 40S initiation complexes obtained in the presence of a ternary complex eIF-2.GTP.Met-tRNA(fMet).

[Affinity modification of the 40S subparticle from human placenta with derivatives of pAUG and pAUGU3].

Affinity labeling of 40S subunits from human placenta with 4-(N-2-chloroethyl-N-methylamino)benzylmethyl-[32P]phosphoamide s of oligoribonucleotides pAUG and pAUGU3 was studied. Covalent attachment of these derivatives to 40S subunits within the complexes with 40S subunits, formed in the presence of Met-tRNAf.eIF-2.

[Affinity modification of 80S ribosomes from human placenta with mRNA analogs--derivatives of oligouridylates with alkylating groups at the 3'-end].

2',3'-O-[4-(N-2-chloroethyl-N-methylamino)]benzylidene derivatives of oligoribonucleotides (Up)5U[32P]pC and (Up)11U[32P]pC were used for affinity labeling of 80S ribosomes from human placenta in the presence of cognate Phe-tRNA(Phe). The derivatives retained the ability of tRNA(Phe)-dependent binding with 80S ribosomes and within the corresponding complexes were cross-linked to 40S subunits (preferentially to 18S rRNA). Sites of the mRNA analogs attachment were identified by blot-hybridization of the modified rRNA with restriction fragments of corresponding rDNA.

[RNA ligase from bacteriophage T4. VIII. Solid phase enzymatic synthesis of oligoribonucleotides].

A method of the solid-phase enzymic synthesis of oligoribonucleotides has been suggested. The donor is fixed through its 3'-end on a water-insoluble matrix followed by the stepwise RNA ligase- and T4 polynucleotide kinase-assisted coupling of trinucleoside diphosphates in the 5'-direction. As an example, (pA)6pAox was immobilised on Biogel P-300 hydrazine and the RNA ligase-catalyzed addition of acceptor ApApA to the donor gave (Ap)9 with the 50% yield.

[RNA-ligase of bacteriophage T4. IV. Synthesis of pentadecaribonucleotide ApUpG(pU)6(pA)5pAp].

Oligoribonucleotide ApUpG(pU)6(pA)5pAp (I) has been prepared by means of RNA ligase. ApUpG, synthesised by the phosphotriester approach, was elongated in the 3'-direction by adding (pU)6 and then (pA)6, which was 3'-blocked with the phosphate or with the periodate-oxidized AMP residue, the latter giving considerably lower level of by-products. Condensation of ApUpG(pU)6 with blocked (pA)6 was carried out under conditions optimal for poor acceptors (20 degrees C, 48 h, pH 8,7) to afford (I) with the yield of 20% (105 OE260); ApUpG(pU)6(pA)10pAp was identified as a byproduct.

[Purification and characteristics of an RNA-ligase preparation from bacteriophage T4].

A purification techniques was developed to obtain a preparation of the bacteriophage T4 RNA-ligase (EC 6.5.1.

[Cytochrome P-450 catalyzed oxidation of 1-piperidinoanthraquinone].

Oxidation of 1-piperidinoantraquinone (1-PA) in microsomal fractions of rat liver was studied. The only product of complete oxidation of 1-PA--(N-antraquinone-1)-delta-aminovaleric acid-was identified using paper and thin-layer chromatography. The participation of cytochrome P-450 in oxidation of 1-PA was demonstrated by sharp inhibition, involving blowing of the microsomes with CO and treatment with sodium deoxycholate.