Exonuclease III
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Exonuclease III
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Identifiers | |
Symbol | ? |
Alt. Symbols | ExoIII |
Entrez | ? |
HUGO | N/A |
OMIM | N/A |
RefSeq | ? |
UniProt | P09030 |
Other data | |
EC number | 3.1.11.2 |
Locus | Chr. N/A [1] |
Exonuclease III (ExoIII) is an enzyme that belongs to the exonuclease family. ExoIII catalyzes the stepwise removal of mononucleotides from 3´-hydroxyl termini of duplex DNA [1]. A limited number of nucleotides are removed during each binding event, resulting in coordinated progressive deletions within the population of DNA molecules [2].
[edit] Function
The preferred substrates are blunt or recessed 3´-termini, although ExoIII also acts at nicks in duplex DNA to produce single-strand gaps. The enzyme is not active on single-stranded DNA, and thus 3´-protruding termini are resistant to cleavage. The degree of resistance depends on the length of the extension, with extensions 4 bases or longer being essentially resistant to cleavage. This property is used to produce unidirectional deletions from a linear molecule with one resistant (3´-overhang) and one susceptible (blunt or 5´-overhang) terminus [3].
ExoIII activity depends partially on the DNA helical structure [4] and displays sequence dependence (C>A=T>G) [5].
ExoIII has also been reported to have RNase H, 3´-phosphatase and AP-endonuclease activities [1].
[edit] Regulation
Temperature, salt concentration and the ratio of enzyme to DNA greatly affect enzyme activity, requiring reaction conditions to be tailored to specific applications.
[edit] References
- Exonuclease III of Escherichia coli K-12, an AP endonuclease. Methods Enzymol. 1980; 65: 201-11.
- Molecular Cloning: A Laboratory Manual. 1989, 2nd. Ed., 5.84-5.85.
- Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984; 28: 351-9 Abstract
- A deoxyribonucleic acid phosphatase-exonuclease from Escherichia coli. II. Characterization of the exonuclease activity. J Biol Chem. 1964; 239: 251-8 Free text (PDF - 937KB)
- Sequence specificity of exonuclease III from E. coli. Nucleic Acids Res. 1982; 10: 4845-59. Free text