scholarly journals In Vitro Ligation of Oligodeoxynucleotides Containing C8-Oxidized Purine Lesions Using Bacteriophage T4 DNA Ligase†

Biochemistry ◽  
2007 ◽  
Vol 46 (12) ◽  
pp. 3734-3744 ◽  
Author(s):  
Xiaobei Zhao ◽  
James G. Muller ◽  
Mohan Halasyam ◽  
Sheila S. David ◽  
Cynthia J. Burrows
Keyword(s):  
Bacteriophage T4 ◽  
Dna Ligase ◽  
T4 Dna Ligase

scholarly journals Effects of DNA-binding drugs on T4 DNA ligase

1990 ◽  
Vol 266 (2) ◽  
pp. 379-384 ◽  
Author(s):  
A Montecucco ◽  
G Pedrali-Noy ◽  
S Spadari ◽  
M Lestingi ◽  
G Ciarrocchi
Keyword(s):  
Dna Binding ◽  
Bacteriophage T4 ◽  
Drug Binding ◽  
Dna Ligase ◽  
Enzyme Complex ◽  
T4 Dna Ligase ◽  
Substrate Interaction ◽  
Dna Nicking

A number of DNA intercalating and externally binding drugs have been found to inhibit nick sealing, cohesive and blunt end ligation, AMP-dependent DNA topoisomerization and EDTA-induced DNA nicking mediated by bacteriophage T4 DNA ligase. The inhibition seems to arise from drug-substrate interaction so that formation of active DNA-Mg2(+)-AMP-enzyme complex is impaired while assembled and active complexes are not disturbed by drug binding to the substrate.

scholarly journals Plant Enzymes but Not AgrobacteriumVirD2 Mediate T-DNA Ligation In Vitro

2000 ◽  
Vol 20 (17) ◽  
pp. 6317-6322 ◽  
Author(s):  
Alicja Ziemienowicz ◽  
Bruno Tinland ◽  
John Bryant ◽  
Veronique Gloeckler ◽  
Barbara Hohn
Keyword(s):  
Plant Extracts ◽  
Test System ◽  
Dna Ligase ◽  
Target Sequence ◽  
T4 Dna Ligase ◽  
Dna Ligases ◽  
Dna Ligation ◽  
Plant Enzymes ◽  
Assay Conditions

ABSTRACT Agrobacterium tumefaciens, a gram-negative soil bacterium, transfers DNA to many plant species. In the plant cell, the transferred DNA (T-DNA) is integrated into the genome. An in vitro ligation-integration assay has been designed to investigate the mechanism of T-DNA ligation and the factors involved in this process. The VirD2 protein, which is produced in Agrobacterium and is covalently attached to T-DNA, did not, under our assay conditions, ligate T-DNA to a model target sequence in vitro. We tested whether plant extracts could ligate T-DNA to target oligonucleotides in our test system. The in vitro ligation-integration reaction did indeed take place in the presence of plant extracts. This reaction was inhibited by dTTP, indicating involvement of a plant DNA ligase. We found that prokaryotic DNA ligases could substitute for plant extracts in this reaction. Ligation of the VirD2-bound oligonucleotide to the target sequence mediated by T4 DNA ligase was less efficient than ligation of a free oligonucleotide to the target. T-DNA ligation mediated by a plant enzyme(s) or T4 DNA ligase requires ATP.

Specificity of the nick-closing activity of bacteriophage T4 DNA ligase

Gene ◽  
1989 ◽  
Vol 76 (2) ◽  
pp. 245-254 ◽  
Author(s):  
Dan Y. Wu ◽  
R.Bruce Wallace
Keyword(s):  
Bacteriophage T4 ◽  
Dna Ligase ◽  
T4 Dna Ligase

Activity-based in vitro selection of T4 DNA ligase

2005 ◽  
Vol 336 (3) ◽  
pp. 987-993 ◽  
Author(s):  
Fumio Takahashi ◽  
Hisakage Funabashi ◽  
Masayasu Mie ◽  
Yaeta Endo ◽  
Tatsuya Sawasaki ◽  
...  
Keyword(s):  
In Vitro Selection ◽  
Dna Ligase ◽  
T4 Dna Ligase ◽  
Selection Of

scholarly journals Homology Modeling of NAD+-Dependent DNA Ligase of the Wolbachia Endosymbiont of Brugia malayi and Its Drug Target Potential Using Dispiro-Cycloalkanones

2015 ◽  
Vol 59 (7) ◽  
pp. 3736-3747 ◽  
Author(s):  
Nidhi Shrivastava ◽  
Jeetendra K. Nag ◽  
Jyoti Pandey ◽  
Rama Pati Tripathi ◽  
Priyanka Shah ◽  
...  
Keyword(s):  
Adverse Effects ◽  
Drug Target ◽  
Brugia Malayi ◽  
Great Promise ◽  
Dna Ligase ◽  
Content Type ◽  
T4 Dna Ligase ◽  
Filarial Nematodes

ABSTRACTLymphatic filarial nematodes maintain a mutualistic relationship with the endosymbiontWolbachia. Depletion ofWolbachiaproduces profound defects in nematode development, fertility, and viability and thus has great promise as a novel approach for treating filarial diseases. NAD+-dependent DNA ligase is an essential enzyme of DNA replication, repair, and recombination. Therefore, in the present study, the antifilarial drug target potential of the NAD+-dependent DNA ligase of theWolbachiasymbiont ofBrugia malayi(wBm-LigA) was investigated using dispiro-cycloalkanone compounds. Dispiro-cycloalkanone specifically inhibited the nick-closing and cohesive-end ligation activities of the enzyme without inhibiting human or T4 DNA ligase. The mode of inhibition was competitive with the NAD+cofactor. Docking studies also revealed the interaction of these compounds with the active site of the target enzyme. The adverse effects of these inhibitors were observed on adult and microfilarial stages ofB. malayiin vitro, and the most active compounds were further monitoredin vivoin jirds and mastomys rodent models. Compounds 1, 2, and 5 had severe adverse effectsin vitroon the motility of both adult worms and microfilariae at low concentrations. Compound 2 was the best inhibitor, with the lowest 50% inhibitory concentration (IC50) (1.02 μM), followed by compound 5 (IC50, 2.3 μM) and compound 1 (IC50, 2.9 μM). These compounds also exhibited the same adverse effect on adult worms and microfilariaein vivo(P< 0.05). These compounds also tremendously reduced the wolbachial load, as evident by quantitative real-time PCR (P< 0.05).wBm-LigA thus shows great promise as an antifilarial drug target, and dispiro-cycloalkanone compounds show great promise as antifilarial lead candidates.

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