Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.7 (DNA polymerase)
 17,007 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The activities of 5-methyltetrahydrofolate (5-CH3-THF)-related enzymes [5-CH3-THF homocysteine methyltransferase and 5,10-methylenetetrahydrofolate (5,10-CH2-THF) reductase] and DNA polymerase alpha were measured in normal and malignant hematopoietic cells. The 5-CH3-THF homocysteine methyltransferase activity was significantly correlated with 5,10-CH2-THF reductase activity, indicating that the hematopoietic cells with active biosynthesis of tetrahydrofolate from 5-CH3-THF also actively synthesize 5-CH3-THF from 5,10-CH2-THF. The activities of 5-CH3-THF-related enzymes had a tendency to be high in lymphoid cells and low in myeloid cells, and were not correlated with the percentage of blasts and immature cells in the samples examined. Fairly good correlations were observed among these three enzymes in non-malignant bone marrow cells. However, the activities of two of the enzymes correlated only weakly overall with DNA polymerase alpha activity in normal and malignant hematopoietic cells. Generally speaking, DNA polymerase alpha activity correlated well with the percentage of blasts and immature cells in the samples examined.
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PMID:5-methyltetrahydrofolate-related enzymes and DNA polymerase alpha in normal and malignant hematopoietic cells. 635 15

Under isothermal conditions, short oligodeoxynucleotides (ODNs) were elongated to long DNA by Vent(exo(-)), a thermophilic DNA polymerase, in the presence of dNTPs. Short ODNs (14-28 nt) were designed to form hairpin structures based on the sequence we obtained from de novo DNA synthesis in the presence of restriction enzyme Tsp509I. As short as 14-nt-long DNA could be elongated to longer than 20000 nucleotides by Vent(exo(-)) at 65 degrees C in 1 h. The high efficiency of elongation at very low concentration (<1 nM) supported the THF-SPE (terminal hairpin formation and self-priming extension) mechanism we purposed for DNA elongation during de novo DNA synthesis. The hairpin structure forms at a DNA duplex end as a self-priming complex, followed by strand displacement extension to longer DNA. The highly efficient elongation attributes to the successive repetition of the process of THF-SPE.
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PMID:Mechanism of DNA elongation during de novo DNA synthesis. 1877 28

A new approach that improves the efficiency and specificity of polymerase chain reaction (PCR) has been developed. Heat-sensitive 3'-protected derivatives of 2'-deoxyribonucleoside 5'-triphosphates (dNTPs) have been synthesized and used as substitutes for natural dTTP, dCTP, dATP, and dGTP in PCR. Since 3'-protected dNTPs are either nonsubstrates or terminating substrates for Taq DNA polymerase, they do not support primer extension/elongation at low stringency conditions during PCR sample preparation when PCR artifacts such as primer dimers and mispriming products can form. At the initial heat-denaturing step and during the PCR sequence, the 3'-protecting group is cleaved, releasing 3'-unprotected dNTP that is a natural substrate for DNA polymerase. As a result, the primer extension/elongation proceeds only at an elevated temperature of PCR, when the interaction of primers and template is highly stringent and specific. Several 3'-protecting groups covering a wide range of deprotection kinetics have been tested. The 3'-O-tetrahydrofuranyl derivatives of dNTPs have demonstrated the best properties leading to a drastically reduced accumulation of PCR artifacts, such as "primer dimers" and "mispriming" products. Overall, PCR with 3'-THF-protected dNTPs demonstrated substantially improved performance and was more efficient and specific compared to PCR with standard dNTPs.
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PMID:3'-Protected 2'-deoxynucleoside 5'-triphosphates as a tool for heat-triggered activation of polymerase chain reaction. 1943 48

N3-methyladenine (3-mA) is a cytotoxic lesion formed by the reaction of DNA with many methylating agents, including antineoplastic drugs, environmental agents and endogenously generated compounds. The toxicity of 3-mA has been attributed to its ability to block DNA polymerization. Using Me-lex, a compound that selectively and efficiently reacts with DNA to afford 3-mA, we have observed in yeast a mutational hotspot at the 5'-terminus of an A(4) tract. In order to explore the potential role of sequence-dependent DNA polymerase bypass of 3-mA, we developed an in vitro system to prepare 3-mA modified substrates using Me-lex. We detail the effects of 3-mA, its stable isostere analogue, 3-methyl-3-deazaadenine, 3-deazaadenine and an THF abasic site on DNA polymerization within an A(4) sequence. The methyl group on 3-mA and 3-methyl-3-deazaadenine has a pronounced inhibitory effect on DNA polymerization. There was no sequence selectivity for the bypass of any of the lesions, except for the abasic site, which was most efficiently by-passed when it was on the 5'-terminus of the A(4) tract. The results indicate that the weak mutational pattern induced by Me-lex may result form the depurination of 3-mA to an abasic site that is bypassed in a sequence dependent context.
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PMID:Effect of n3-methyladenine and an isosteric stable analogue on DNA polymerization. 2093 69