RESUMO
1,2-Diol-oligoribonucleotides were prepared using fully protected 2'-O-[2-(2,3-dihydroxypropyl)amino-2-oxoethyl]uridine 3'-phosphoramidite. Incorporation of the 2'-modified uridine residue into oligonucleotide chains does not significantly affect the thermal stability of RNA and RNA-DNA duplexes. Periodate oxidation of the 1,2-diol results in reactive 2'-aldehyde oligoribonucleotides. Further application of these oligonucleotides for cross-linking with bacterial ribonuclease P was investigated.
Assuntos
Aldeídos/química , DNA/química , Ácidos Nucleicos Heteroduplexes/química , Oligorribonucleotídeos/química , Oligorribonucleotídeos/síntese química , RNA/química , Proteínas de Bactérias/química , Ribonuclease P/químicaRESUMO
Oligonucleotides with 1,2-diol grouping were prepared from 2'-O-[2-(2,3-dihydroxypropyl)amino-2-oxo-ethyl]uridine 3'-phosphoramidite. The thermal stability of modified DNA duplexes and their ability to form complexes with the p50 subunit of the NF-kappaB transcription factor and (cytosine-5)-DNA methyltransferase SsoII were studied. The periodate oxidation of the l,2-diol grouping of the oligonucleotides resulted in reactive 2'-aldehyde derivatives. The opportunity of their use for the affinity modification of DNA-recognizing proteins was studied.
Assuntos
Aldeídos/síntese química , DNA-Citosina Metilases/química , Subunidade p50 de NF-kappa B/química , Oligonucleotídeos/síntese química , Uridina/análogos & derivados , Uridina/síntese química , Aldeídos/química , Ligantes , Oligonucleotídeos/química , Uridina/químicaRESUMO
Integration of human immunodeficiency virus type 1 DNA into the infected cell genome is one of the key steps of the viral replication cycle. Therefore viral enzyme integrase, which realizes the integration, is of interest as a target for new antiviral drugs. Conjugates of 11-mer single stranded oligonucleotides with hydrophobic molecules are shown to be efficient integrase inhibitors since they induce dissociation of the integrase-viral DNA complex. The effect of the oligonucleotide length and structure as well as the structure of hydrophobic molecules on the conjugate inhibitory activity has been studied. Conjugates with eosin and oleic acid are shown to be the most active. Conjugates of these molecules with 2'-O-methyl-oligonucleotide inhibit integrase at 50-100 nM and have no influence on a number of other DNA-binding enzymes.
Assuntos
DNA Viral/química , Inibidores Enzimáticos/química , Amarelo de Eosina-(YS)/química , Integrase de HIV/química , Ácido Oleico/química , Oligonucleotídeos/química , Proteínas de Ligação a DNA/química , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Genoma Humano , Humanos , Oligonucleotídeos/farmacologiaRESUMO
DNA duplexes containing the iodoacetamido group at position 2' of the ribose moiety were proposed for affinity modification of Cys in DNA-binding proteins. Reactive DNA derivatives were obtained with iodoacetic anhydride and synthetic oligodeoxyribonucleotides containing 2'-amino-2'-deoxyuridine in place of thymine at various positions. The derivatives were tested for reaction with amino acids and peptides and shown to specifically interact with Cys-containing proteins. The possibility of using the modified DNA duplexes to probe the protein SH group close to the DNA sugar-phosphate backbone in DNA-protein complexes was demonstrated with the example of subunit p50 of human transcription factor NF-kappa B.
Assuntos
DNA/química , Indicadores e Reagentes/química , Iodoacetatos/química , Ácidos Nucleicos Heteroduplexes/química , Proteínas/química , Nucleotídeos de Uracila/química , Cisteína/química , Dimerização , Glutationa Transferase/química , Glutationa Transferase/genética , Humanos , NF-kappa B/química , NF-kappa B/genética , Subunidade p50 de NF-kappa B , Oligonucleotídeos/química , Ribose/químicaRESUMO
Oligonucleotide-peptide conjugates have several applications, including their potential use as improved antisense agents for interfering with the RNA function within cells. In order to provide robust and generally applicable conjugation chemistry, we developed a novel approach of fragment coupling of pre-synthesized peptides to the 2'-position of a selected nucleotide within an otherwise protected oligonucleotide chain attached to a solid support.
Assuntos
Bioquímica/métodos , Nucleoproteínas/química , Oligonucleotídeos Antissenso/química , Oligonucleotídeos/química , Peptídeos/química , Adenosina/análogos & derivados , Adenosina/química , Arabinonucleosídeos/química , Cromatografia Líquida de Alta Pressão , Nucleoproteínas/síntese química , Oligonucleotídeos/síntese química , Oligonucleotídeos Antissenso/síntese química , Peptídeos/síntese química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
A method for preparing a modified derivative of 2'-amino-2'-deoxy-arabino-adenosine ready for directed insertion into an oligonucleotide chain during solid-phase synthesis was elaborated. A series of the title oligonucleotides (6-25-mers) containing a 2'-amino-2'-deoxy-arabino-adenosine fragment were prepared. A high reactivity of the 2'-amino group during the acylation with carboxylic acid anhydrides was demonstrated. It was shown that the insertion of the modified fragments into oligonucleotides did not inhibit the formation of the DNA duplex.
