The metastasis suppressor NM23-H1 possesses 3'-5' exonuclease activity

Deqin Ma; Joseph R McCorkle; David M Kaetzel

doi:10.1074/jbc.M400185200

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The metastasis suppressor NM23-H1 possesses 3'-5' exonuclease activity

Journal article

Open access

Peer reviewed

The metastasis suppressor NM23-H1 possesses 3'-5' exonuclease activity

Deqin Ma, Joseph R McCorkle and David M Kaetzel

The Journal of biological chemistry, Vol.279(17), pp.18073-18084

04/23/2004

DOI: 10.1074/jbc.M400185200

PMID: 14960567

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Abstract

NM23-H1 belongs to a family of eight gene products in humans that have been implicated in cellular differentiation and development, as well as oncogenesis and tumor metastasis. We have defined NM23-H1 biochemically as a 3'-5' exonuclease by virtue of its ability in stoichiometric amounts to excise single nucleotides in a stepwise manner from the 3' terminus of DNA. The activity is dependent upon the presence of Mg(2+), is most pronounced with single-stranded substrates or mismatched bases at the 3' terminus of double-stranded substrates, and is inhibited by both ATP and the incorporation of cordycepin, a 2'-deoxyadenosine analogue, into the 3'-terminal position. The 3'-5' exonuclease activity was assigned to NM23-H1 by virtue of: 1) precise coelution of enzymatic activity with wild-type and mutant forms of NM23-H1 protein during purification by hydroxylapatite and gel filtration column high performance liquid chromatography and 2) significantly diminished activity exhibited by purified recombinant mutant forms of the proteins. Lysine 12 appears to play an important role in the catalytic mechanism, as evidenced by the significant reduction in 3'-5' exonuclease activity resulting from a Lys(12) to glutamine substitution within the protein. 3'-5' Exonucleases are believed to play an important role in DNA repair, a logical candidate function underlying the putative antimetastatic and oncogenic activities of NM23-H1.

Protein Structure, Tertiary

Proteins - physiology

Deoxyadenosines - chemistry

Mutagenesis, Site-Directed

Humans

Oligodeoxyribonucleotides - chemistry

Recombinant Proteins - chemistry

Chromatography, High Pressure Liquid

DNA - chemistry

Exonucleases - chemistry

Magnesium - chemistry

Neoplasm Metastasis

Nucleoside-Diphosphate Kinase - chemistry

Proteins - metabolism

Time Factors

DNA Repair

NM23 Nucleoside Diphosphate Kinases

Models, Genetic

Mutation

Adenosine Triphosphate - chemistry

Chromatography, Gel

Lysine - chemistry

Circular Dichroism

Details

Title: Subtitle: The metastasis suppressor NM23-H1 possesses 3'-5' exonuclease activity
Creators: Deqin Ma - Department of Molecular and Biomedical Pharmacology, University of Kentucky Medical Center, Lexington, Kentucky 40536-0084, USA
Joseph R McCorkle
David M Kaetzel
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.279(17), pp.18073-18084
DOI: 10.1074/jbc.M400185200
PMID: 14960567
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: CA83237 / NCI NIH HHS
Language: English
Date published: 04/23/2004
Academic Unit: Pathology
Record Identifier: 9984047693602771

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