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PMID: 8811082 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Molecular cloning and functional analysis of a Schizosaccharomyces pombe homologue of Escherichia coli endonuclease III.

Nucleic acids research ·Vol. 24 ·No. 17 ·1996-09-01 ·Pages 3307-12

Roldán-Arjona T, Anselmino C, Lindahl T

Abstract

The Escherichia coli endonuclease III (Nth-Eco) protein is involved in the removal of damaged pyrimidine residues from DNA by base excision repair. It is an iron-sulphur enzyme possessing both DNA glycosylase and apurinic/apyrimidinic lyase activities. A database homology search identified an open reading frame in genomic sequences of Schizosaccharomyces pombe which encodes a protein highly similar to Nth-Eco. The gene has been subcloned in an expression vector and the protein purified to apparent homogeneity. The S.pombe Nth homologue (Nth-Spo) is a 40.2 kDa protein of 355 amino acids. Nth-Spo possesses glycosylase activity on different types of DNA substrates with pyrimidine damage, being able to release both urea and thymine glycol from double-stranded polymers. The eukaryotic protein removes urea more efficiently than the prokaryotic enzyme, whereas its efficiency in excising thymine glycol is lower. A nicking assay was used to show that the enzyme also exhibits an AP lyase activity on UV- and gamma-irradiated DNA substrates. These findings show that Nth protein is structurally and functionally conserved from bacteria to fission yeast.

MeSH Terms
Amino Acid Sequence Cloning, Molecular DNA Glycosylases DNA Repair DNA-(Apurinic or Apyrimidinic Site) Lyase Deoxyribonuclease (Pyrimidine Dimer) Deoxyribonuclease IV (Phage T4-Induced) Endodeoxyribonucleases/genetics,metabolism Escherichia coli Proteins Eukaryotic Cells Lyases/analysis Molecular Sequence Data N-Glycosyl Hydrolases/analysis Recombinant Proteins/metabolism Schizosaccharomyces/enzymology,genetics Sequence Analysis, DNA Sequence Homology, Amino Acid Thymine/analogs & derivatives,metabolism Urea/metabolism
Chemicals
Escherichia coli Proteins Recombinant Proteins thymine glycol Urea Endodeoxyribonucleases Deoxyribonuclease IV (Phage T4-Induced) endonuclease IV, E coli Deoxyribonuclease (Pyrimidine Dimer) NTH protein, E coli DNA Glycosylases N-Glycosyl Hydrolases Lyases DNA-(Apurinic or Apyrimidinic Site) Lyase Thymine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Roldán-Arjona T
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Hertfordshire, UK.
Anselmino C
Lindahl T
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42 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/24.17.3307
Published
1996-09-01
Pages
3307-12
Language
English
Region
England
NLM ID
0411011
PMCID
PMC146095
Subset
IM
Databases
SWISSPROT
P31378
External Links
PubMed source DOI: 10.1093/nar/24.17.3307 PMC Full Text View journal details
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