Deoxyribonucleic Acid Ligase Research Articles

Role for deoxyribonucleic acid ligase in deoxyribonucleic acid polymerase i-dependent repair synthesis in toluene-treated escherichia coli

In a toluene-treated mutant of Escherichia coli K-12 having a temperature-sensitive, conditionally lethal mutation in the structural gene for deoxyribonucleic acid (DNA) ligase, an extensive DNA repair synthesis occurred in X-irradiated cells at the nonpermissive temperature, 42 C. At the permissive temperature, 30 C, nearly normal semiconservative synthesis and limited repair synthesis were observed when DNA ligase was activated by the addition of nicotinamide adenine dinucleotide.

A deoxyribonucleic acid ligase from nuclei of rat liver. Purification and properties.

A deoxyribonucleic acid ligase from nuclei of rat liver. Purification and properties.

Ribonucleic acid ligase activity of deoxyribonucleic acid ligase from phage T4 infected Escherichia coli.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTRibonucleic acid ligase activity of deoxyribonucleic acid ligase from phage T4 infected Escherichia coliHiroshi Sano and Guenter FeixCite this: Biochemistry 1974, 13, 25, 5110–5115Publication Date (Print):December 1, 1974Publication History Published online1 May 2002Published inissue 1 December 1974https://doi.org/10.1021/bi00722a009RIGHTS & PERMISSIONSArticle Views55Altmetric-Citations14LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (632 KB) Get e-Alerts Get e-Alerts

Covalent Attachment of Polyribonucleotides to Polydeoxyribonucleotides Catalyzed by Deoxyribonucleic Acid Ligase

Abstract DNA ligase isolated from Escherichia coli or phage T4-infected E. coli catalyzes the covalent joining of the 5'-phosphate termini of polydeoxyribonucleotides and 3'-hydroxyl termini of polyribonucleotides. This reaction occurs with poly(dA) and poly(A) in the presence of poly(dT). The 5'-phosphate terminus of poly[d(A-T)] can be linked by an intramolecular reaction to its 3'-hydroxyl terminus substituted with a ribonucleotide. Phage T4-induced DNA ligase also joins poly(dT) to poly(U) in the presence of poly(dA). The joining of 5'-phosphate termini of polyribonucleotides to 3'-hydroxyl termini of polydeoxyribonucleotides was detected only with phage T4-induced DNA ligase. This reaction, however, occurred to a limited extent with poly(dA) substrates containing AMP residues at the 5' terminus in the presence of poly(dT). These results suggest that although DNA ligase from E. coli is incapable of joining RNA to RNA, it is capable of joining 5'-phosphate terminus of a DNA to the 3'-hydroxyl terminus of RNA in addition to the well known joining reaction between DNA to DNA. The phage T4-induced DNA ligase on the other hand joins DNA to DNA, RNA to RNA, and RNA to DNA in all combinations.

In situ activity of enzymes on polyacrylamide gels of a deoxyribonucleic acid-membrane fraction extracted from pneumococci.

A deoxyribonucleic acid (DNA)-membrane fraction extracted from Diplococcus pneumoniae was subjected to polyacrylamide gel electrophoresis after treatment with 0.16% sodium dodecyl sulfate. At least two DNA polymerase activities were detected by in situ assays with appropriate substrates, templates, and inhibitors, including a co-polymer of deoxyadenylic and thymidylic acid and N-ethylmaleimide. This activity coincided with a fraction in the gel containing 7.5, 9.4, and 24%, respectively of the DNA, phospholipid, and protein present in the DNA-membrane fraction before electrophoresis and sodium dodecyl sulfate treatment. Assays with minced gels showed that several nuclease activities, deoxyribonucleotide kinase activity, and DNA ligase activity also coincided with this fraction. However, ribonucleoside diphosphate reductase activity did not. These results demonstrate that a complex of enzymes involved in DNA replication is firmly bound to the DNA-membrane fraction in pneumococci.

Ligase-defective bacteriophage T4. II. Physiological studies.

The timing of the suppression of gene 30 (deoxyribonucleic acid ligase) mutations by rII mutations was studied by temperature shift-down experiments with a temperature-sensitive rII mutation. The rII function must remain inactivated for about 5 to 8 min at 37 C for suppression to occur, thus making suppression an early function. This result is in agreement with the timing of expression of other rII functions. A gene 30 defect can also be overcome by replacing the Na(+) cation in the growth medium with the Mg(2+) cation, a result similar to the relief of the lethality of rII mutations in lambda lysogens. Prior infection with bacteriophages T3 or T7, which produce their own deoxyribonucleic acid ligases, can also partially overcome the lethality of gene 30 mutations.

Ligase-defective bacteriophage T4. I. Effects on mutation rates.

Temperature-sensitive mutations in bacteriophage T4 gene 30 (polynucleotide ligase) were examined for their effects on spontaneous and proflavine-induced frameshift mutagenesis in the rII and ac (acridine resistance) cistrons. Only small (fourfold or less) effects on mutation rates were observed, even when selection artifacts involving suppression of gene 30 mutations by rII mutations were taken into account. The deoxyribonucleic acid ligase gene of T4 therefore appears to be only a minor determinant of frameshift mutation rates. This result is consistent with the particular nature of frameshift mutagenesis in bacteriophage T4.

Mutagen stability of alkylation-sensitive mutants of Bacillus subtilis.

A series of mutations of Bacillus subtilis, conferring sensitivity to methyl methanesulfonate (MMS), were transferred by transformation to a suppressible his(-) stock. The introduction of certain sensitivity mutations prevented the ultraviolet- or MMS-induced, but not the spontaneous, reversion of his(-) to his(+). Not all sensitivity mutations led to this resistance to mutagenesis; a strain with altered deoxyribonucleic acid (DNA) polymerase activity behaved almost normally with respect to its mutagen response, as did an excision-defective, ultraviolet-sensitive strain used as a control. One of the mutagen-stable strains responded to mutagenesis with nitrosomethylguanidine; another appeared stable even to this mutagen. All mutagen-stable strains had DNA polymerase and DNA ligase activity.

Inactivation of bacteriophage T4 by ethyl methanesulfonate: influence of host and viral genotypes.

Inactivation of bacteriophage T4 by ethyl methanesulfonate (EMS) is a complex process which depends critically upon the conditions of treatment and upon both the viral and the host genotypes. EMS-inactivated particles are capable of multiplicity and cross-reactivation, indicating the need for caution in using EMS in certain types of mutation studies. The pyrimidine dimer excision systems of the phage and the host do not affect the EMS sensitivity of T4, but the T4x(+)y(+) system does. Mutational defects in the deoxyribonucleic acid (DNA) ligase and the DNA polymerase systems both of the virus and of its host also affect viral EMS sensitivity.

Influence of thymine starvation on the integrity of deoxyribonucleic acid in Escherichia coli.

The influence of thymine starvation on the single-strand molecular weight of deoxyribonucleic acid (DNA) from Escherichia coli was determined by sedimentation through gradients of alkaline sucrose. Growth of cells for as long as 150 min in thymineless medium did not significantly reduce the molecular weight below the control value of 2.4 +/- 0.3 x 10(8) daltons. Incubation of cells in thymineless medium after exposure to 500 ergs/mm(2) of ultraviolet light or 20 krad of (137)Cs gamma rays did not appear to block the rejoining of single-strand breaks associated with irradiation. Thus, DNA repair enzymes, presumably including DNA ligase, are not significantly inhibited by thymine starvation.

Properties of Bacteriophage T4 Mutants Defective in Gene 30 (Deoxyribonucleic Acid Ligase) and the rII Gene

In Escherichia coli K-12 strains infected with phage T4 which is defective in gene 30 [deoxyribonucleic acid (DNA) ligase] and in the rII gene (product unknown), near normal levels of DNA and viable phage were produced. Growth of such T4 ligase-rII double mutants was less efficient in E. coli B strains which show the "rapidlysis" phenotype of rII mutations. In pulse-chase experiments coupled with temperature shifts and with inhibition of DNA synthesis, it was observed that DNA synthesized by gene 30-defective phage is more susceptible to breakdown in vivo when the phage is carrying a wild-type rII gene. Breakdown was delayed or inhibited by continued DNA synthesis. Mutations of the rII gene decreased but did not completely abolish the breakdown. T4 ligase-rII double mutants had normal sensitivity to ultraviolet irradiation.

Properties of a deoxyribonucleic acid ligase mutant of Escherichia coli: x-ray sensitivity.

A deoxyribonucleic acid ligase-deficient mutant is X-ray sensitive relative to the parent strain, suggesting that deoxyribonucleic acid ligase functions in repair of X-ray-induced, single-strand scissions.

Enzymatic Joining of Deoxyribonucleic Acid Strands: III. Further purification of the deoxyribonucleic acid ligase from escherichia coli and multiple forms of the purified enzyme

Abstract The DNA ligase from Escherichia coli forms an intermediate enzyme-bound complex with the AMP moiety of DPN+ (ligase-AMP). Detection of this complex was the basis for a simple new assay used in a revised, more extensive purification of the enzyme. Ligase-AMP made from the most highly purified enzyme fraction readily converts to two new chromatographically distinguishable forms (I and II) which differ from the original form (Form III) in their functional capacities. Only Form III is active in the routine DNA joining assay previously described. All three forms discharge their AMP residue in reaction with nicotinamide mononucleotide. Form I does not discharge its AMP residue with DNA bearing single strand interruptions, whereas Form II does so only with much higher DNA concentrations than those that discharge the third form. Gel filtration suggests that Form I has a molecular weight of 5 x 104 daltons compared with 10 x 104 daltons for the other forms.