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Ann N Y Acad Sci ; 1091: 319-35, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17341625


The role of various inflammatory mechanisms and oxidative stress in the development of atherosclerosis and arterial hypertension (AH) has been increasingly acknowledged during recent years. Hypertension per se or factors that cause hypertension along with other complications lead to infiltration of activated leukocytes in the vascular wall, where these cells contribute to the development of vascular injury by releasing cytokines, oxygen radicals, and other toxic mediators. However, molecular mechanisms underlying leukocyte activation at transcriptional level in AH are still far from being clear. To solve this problem we employed cDNA microarray technology to reveal the differences in gene expression in peripheral blood leukocytes from patients with AH compared with healthy individuals. The microarray data were verified by a semi-quantitative RT-PCR method. We found 25 genes with differential expression in leukocytes from AH patients among which 21 genes were upregulated and 4 genes were downregulated. These genes are implicated in apoptosis (CASP2, CASP4, and CASP8, p53, UBID4, NAT1, and Fte-1), inflammatory response (CAGC, CXCR4, and CX3CR1), control of MAP kinase function (PYST1, PAC1, RAF1, and RAFB1), vesicular trafficking of molecules among cellular organelles (GDI-1 and GDI-2), cell redox homeostasis (GLRX), cellular stress (HSPA8 and HSP40), and other processes. Gene expression pattern of the majority of genes was similar in AH patients independent of the disease stage and used hypotensive therapy, but was clearly different from that of normotensive subjects.

Perfilação da Expressão Gênica , Regulação da Expressão Gênica/fisiologia , Hipertensão/genética , Hipertensão/metabolismo , Leucócitos/metabolismo , Adulto , Idoso , Feminino , Humanos , Hipertensão/patologia , Masculino , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos
Biochim Biophys Acta ; 1008(1): 113-5, 1989 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-2497780


The ability of a wide variety of nucleoside 5'-triphosphates with modified sugar moiety to serve as substrates in DNA synthesis catalyzed by DNA polymerase A from the archaebacterium Sulfolobus acidocaldarius was studied. Most of the dNTP analogs tested are shown to be specific terminating substrates for the synthesis irreversibly blocking further elongation of a nascent chain. The most powerful inhibitors were found to be 3'-amino derivatives of deoxy and arabino nucleoside triphosphates, while specific reverse transcriptase inhibitors, 3'-azido and 3'-methoxy derivatives of dNTP, were found to be inactive.

Archaea/enzimologia , Bactérias/enzimologia , DNA Polimerase Dirigida por DNA/metabolismo , Desoxirribonucleotídeos/metabolismo , Sequência de Bases , DNA Bacteriano/biossíntese , Dados de Sequência Molecular , Especificidade por Substrato
FEBS Lett ; 228(1): 144-8, 1988 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-2449362


The influence of a double-stranded region on DNA synthesis performed by a series of DNA polymerases on a single-stranded template was studied. Two types of double-stranded hindrances were employed: a stable hairpin formed by the template alone and a region formed by the template and an extraneous oligonucleotide complementary to the template. While T4 and calf thymus alpha DNA polymerases are strongly arrested at the beginning of either of the two double-stranded hindrances, the Klenow fragment of E. coli DNA polymerase I and avian myeloblastosis virus reverse transcriptase can pass through the double-stranded regions. The DNA chain-elongation rate seems to be undisturbed in the case of reverse transcriptase but greatly reduced for DNA polymerase I.

DNA Polimerase II/metabolismo , DNA Polimerase I/metabolismo , DNA Polimerase Dirigida por DNA/metabolismo , DNA/biossíntese , DNA Polimerase Dirigida por RNA/metabolismo , Animais , Vírus da Mieloblastose Aviária/enzimologia , Bovinos , Escherichia coli/enzimologia , Relação Estrutura-Atividade , Fagos T/enzimologia , Timo/enzimologia
Biochim Biophys Acta ; 868(2-3): 136-44, 1986 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-2429703


Terminal deoxynucleotidyl transferase from calf thymus and RNA-directed DNA polymerase (reverse transcriptase) from the avian myeloblastosis virus catalyze the incorporation of 3'-amino-2',3'-dideoxynucleoside 5'-triphosphates, as well as some of their 3'-derivatives, 3'-amino-3'-deoxyarabinonucleoside 5'-triphosphates and some other nucleoside 5'-triphosphates modified at sugar residues. After incorporation of the appropriate 5'-mononucleotide residue into the DNA, further chain elongation is blocked. This finding opens up a possibility for selective inhibition of DNA synthesis catalyzed by a certain enzyme.

DNA Nucleotidilexotransferase/metabolismo , DNA Nucleotidiltransferases/metabolismo , Nucleotídeos/metabolismo , DNA Polimerase Dirigida por RNA/metabolismo , Animais , Vírus da Mieloblastose Aviária/enzimologia , Catálise , Bovinos , DNA/biossíntese , DNA Nucleotidilexotransferase/antagonistas & inibidores , DNA de Cadeia Simples/metabolismo , Nucleotídeos/farmacologia , Inibidores da Transcriptase Reversa , Especificidade por Substrato , Moldes Genéticos
FEBS Lett ; 183(2): 275-8, 1985 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-2580738


It is shown that dNTP(3'F) are terminators of DNA synthesis and may serve as very effective tools for DNA sequencing with E.coli DNA polymerase I and AMV reverse transcriptase. The dNTP(3'F) are found to be chain terminator substrates for calf thymus terminal deoxyribonucleotidyl transferase but not for calf thymus DNA polymerase alpha. The optimal dNTP(3'F) concentration for DNA sequencing by DNA polymerase I is found to be an order of magnitude lower than that of ddNTPs. dNTP(3'F) produce a more clear sequence pattern than do ddNTPs.

Replicação do DNA/efeitos dos fármacos , Desoxirribonucleotídeos/farmacologia , Flúor/farmacologia , Animais , Sequência de Bases , Bovinos , DNA Nucleotidilexotransferase/metabolismo , DNA Polimerase I/metabolismo , Escherichia coli , DNA Polimerase Dirigida por RNA/metabolismo
FEBS Lett ; 164(1): 97-101, 1983 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-6228440


We have analyzed the specificity of the actinomycin D-DNA interaction. The 'footprint' method has been used in this investigation. It is shown that: (i) The presence of dinucleotide GC or GG is required for binding of a single drug molecule. (ii) The strong binding sites are encoded by tetranucleotide XGCY; where X not equal to G and Y not equal to C in accordance with RNA elongation hindrance sites [1]. (iii) There is a positive cooperativity in binding of actinomycin D with DNA.

DNA , Dactinomicina , Endodesoxirribonucleases/metabolismo , Sequência de Bases , Fenômenos Químicos , Química , Desoxirribonuclease I , Plasmídeos , Especificidade por Substrato