Characterization of a Baculovirus Enzyme with RNA Ligase, Polynucleotide 5′-Kinase, and Polynucleotide 3′-Phosphatase Activities
نویسندگان
چکیده
منابع مشابه
Induced Polynucleotide Kinase, Polynucleotide Ligase, and Deoxyribonucleic Acid Polymeraset
The mechanism of action of T4-induced polynu-cleotide kinase on short single-stranded deoxyoligonucleotideshas been investigated.It was found that the phosphorylationreaction catalyzed by T, polynucleotide kinase could be re-versed. Thus, a 5 ’-3*P-labeleddeoxyoligonucleotide and ADPon incubation with the enzyme formed [Y-~~PJATP.In addi-tion some radioac...
متن کاملCharacterization of the 2′,3′ cyclic phosphodiesterase activities of Clostridium thermocellum polynucleotide kinase-phosphatase and bacteriophage λ phosphatase
Clostridium thermocellum polynucleotide kinase-phosphatase (CthPnkp) catalyzes 5' and 3' end-healing reactions that prepare broken RNA termini for sealing by RNA ligase. The central phosphatase domain of CthPnkp belongs to the dinuclear metallophosphoesterase superfamily exemplified by bacteriophage lambda phosphatase (lambda-Pase). CthPnkp is a Ni(2+)/Mn(2+)-dependent phosphodiesterase-monoest...
متن کاملMutational analysis defines the 5'-kinase and 3'-phosphatase active sites of T4 polynucleotide kinase.
T4 polynucleotide kinase (Pnk) is a bifunctional 5'-kinase/3'-phosphatase that aids in the repair of broken termini in RNA by converting 3'-PO4/5'-OH ends into 3'-OH/5'-PO4 ends, which are then sealed by RNA ligase. Here we have employed site-directed mutagenesis (introducing 31 mutations at 16 positions) to locate candidate catalytic residues within the 301 amino acid Pnk polypeptide. We found...
متن کاملDNA end processing by polynucleotide kinase/phosphatase.
S cission of one or both strands of the DNA duplex occurs as ionizing radiation and oxidative DNA damage mount a constant assault our genomes. Like a broken limb, or split piece of wood, DNA strand breaks are typically not clean. That is, they lack the DNA 3′-hydroxyl and 5′-phosphate moieties needed for DNA repair synthesis by DNA polymerases and DNA end joining by DNA ligases (Fig. 1A). For i...
متن کاملMechanism of RNA 2′,3′-cyclic phosphate end healing by T4 polynucleotide kinase–phosphatase
T4 polynucleotide kinase-phosphatase (Pnkp) exemplifies a family of enzymes with 5'-kinase and 3'-phosphatase activities that function in nucleic acid repair. The polynucleotide 3'-phosphatase reaction is executed by the Pnkp C-terminal domain, which belongs to the DxDxT acylphosphatase superfamily. The 3'-phosphatase reaction entails formation and hydrolysis of a covalent enzyme-(Asp165)-phosp...
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ژورنال
عنوان ژورنال: Journal of Biological Chemistry
سال: 2004
ISSN: 0021-9258
DOI: 10.1074/jbc.m313386200