Exonuclease III explained

Exonuclease III
Organism:E. coli strain K-12/MG1655
Taxid:511145
Symbol:xthA
Altsymbols:ExoIII
Entrezgene:946254
Refseqprotein:NP_416263
Uniprot:P09030
Ecnumber:3.1.11.2
Chromosome:genome
Entrezchromosome:NC_000913
Genloc Start:1830330
Genloc End:1831378

Exonuclease III (ExoIII) is an enzyme that belongs to the exonuclease family. ExoIII catalyzes the stepwise removal of mononucleotides from 3´-hydroxyl termini of double-stranded DNA. A limited number of nucleotides are removed during each binding event, resulting in coordinated progressive deletions within the population of DNA molecules.

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.[1]

ExoIII activity depends partially on the DNA helical structure[2] and displays sequence dependence (C>A=T>G).[3]

ExoIII has also been reported to have RNase H, 3´-phosphatase and AP-endonuclease activities.

Current Studies

There are many different exonucleases and many are still to be discovered in bacteria, current studies are being conducted in E. coli. Many exonucleases fall into superfamilies with different domains of life proving that exonuclease III has shown to be ancient. Exonucleases evolved early in the history of life and have vital biological roles.[4]

Regulation

Temperature, salt concentration and the ratio of enzyme to DNA greatly affect enzyme activity, requiring reaction conditions to be tailored to specific applications.

Further reading

Notes and References

  1. Book: Rogers SG, Weiss B . Exonuclease III of Escherichia coli K-12, an AP endonuclease . Nucleic Acids Part I . 65 . 1 . 201–11 . 1980 . 6246343 . 10.1016/S0076-6879(80)65028-9 . 978-0-12-181965-1 . Methods in Enzymology . https://archive.org/details/nucleicacids0000unse/page/201 .
  2. Book: Maniatis . Tom . Sambrook . Joseph . Fritsch EF . vanc . Molecular cloning: a laboratory manual . Cold Spring Harbor Laboratory . Cold Spring Harbor, N.Y . 1989 . 5.84–5 . 978-0-87969-309-1 . 2nd .
  3. Henikoff S . Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing . Gene . 28 . 3 . 351–9 . June 1984 . 6235151 . 10.1016/0378-1119(84)90153-7 .
  4. Lovett ST . The DNA Exonucleases of Escherichia coli . EcoSal Plus . 4 . 2 . December 2011 . 26442508 . 4238392 . 10.1128/ecosalplus.4.4.7 .