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Titolo:
Functional domains of an NAD(+)-dependent DNA ligase
Autore:
Timson, DJ; Wigley, DB;
Indirizzi:
Univ Oxford, Sir William Dunn Sch Pathol, Oxford OX1 3RE, England Univ Oxford Oxford England OX1 3RE n Sch Pathol, Oxford OX1 3RE, England
Titolo Testata:
JOURNAL OF MOLECULAR BIOLOGY
fascicolo: 1, volume: 285, anno: 1999,
pagine: 73 - 83
SICI:
0022-2836(19990108)285:1<73:FDOAND>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
RNA CAPPING ENZYME; POLY(ADP-RIBOSE) POLYMERASE; COVALENT CATALYSIS; ESCHERICHIA-COLI; BACTERIOPHAGE-T7; SUPERFAMILY; MUTAGENESIS; PROTEINS;
Keywords:
DNA replication; limited proteolysis; DNA binding; protein-DNA recognition; zinc binding protein;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
31
Recensione:
Indirizzi per estratti:
Indirizzo: Wigley, DB Univndxford, Sir William Dunn Sch Pathol, S Parks Rd, Oxford OX1 3RE, Engla Univ Oxford S Parks Rd Oxford England OX1 3RE d OX1 3RE, Engla
Citazione:
D.J. Timson e D.B. Wigley, "Functional domains of an NAD(+)-dependent DNA ligase", J MOL BIOL, 285(1), 1999, pp. 73-83

Abstract

Limited proteolysis of the NAD(+)-dependent DNA ligase from Bacillus stearothermophilus with thermolysin results in two fragments which were resistant to further proteolysis. These fragments were characterised by N-terminal protein sequencing and electrospray mass spectrometry. The larger, N-terminal fragment consists of the first 318 residues and the smaller, C-terminal fragment begins at residue 397 and runs to the C terminus. Both fragments were over-expressed in Escherichia coli and purified to homogeneity from this source. The large fragment retains the full self-adenylation activity of the intact enzyme, has minimal DNA binding activity and vastly reduced Ligation activity. The small fragment lacks adenylation activity but binds to nicked DNA with a similar affinity to that of the intact enzyme. It is unable to stimulate the ligation activity of the large fragment. Atomic absorption spectroscopy showed that the intact protein and the small fragment bind a zinc ion but the large fragment does not. No evidence of any interaction between the two fragments could be obtained. Thus, we conclude that NAD+-dependent DNA ligases consist of at least two discrete functional domains: anN-terminal domain which is responsible for cofactor binding and self adenylation, and a C-terminal DNA-binding domain which contains a zinc binding site. (C) 1999 Academic Press.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 21/09/20 alle ore 15:30:45