Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (422K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (4)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Jokela, M.
Right arrow Articles by Syväoja, J. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jokela, M.
Right arrow Articles by Syväoja, J. E.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 30 April 2004

Nucleic Acids Research, 2004, Vol. 32, No. 8 2430-2440
© 2004 Oxford University Press

Characterization of the 3' exonuclease subunit DP1 of Methanococcus jannaschii replicative DNA polymerase D

Maarit Jokela1, Anitta Eskelinen1, Helmut Pospiech1, Juha Rouvinen2 and Juhani E. Syväoja*,3

1 Biocenter Oulu and Department of Biochemistry, PO Box 3000, FIN-90014 University of Oulu, Finland, 2 Department of Chemistry and 3 Department of Biology, University of Joensuu, PO Box 111, FIN-80101 Joensuu, Finland

*To whom correspondence should be addressed. Tel: +358 13 251 3697; Fax: +358 13 251 3590; Email: juhani.syvaoja{at}joensuu.fi

Received February 4, 2004; Revised March 11, 2004; Accepted April 1, 2004

The B-subunits associated with the replicative DNA polymerases are conserved from Archaea to humans, whereas the corresponding catalytic subunits are not related. The latter belong to the B and D DNA polymerase families in eukaryotes and archaea, respectively. Sequence analysis places the B-subunits within the calcineurin-like phosphoesterase superfamily. Since residues implicated in metal binding and catalysis are well conserved in archaeal family D DNA polymerases, it has been hypothesized that the B-subunit could be responsible for the 3'-5' proofreading exonuclease activity of these enzymes. To test this hypothesis we expressed Methanococcus jannaschii DP1 (MjaDP1), the B-subunit of DNA polymerase D, in Escherichia coli, and demonstrate that MjaDP1 functions alone as a moderately active, thermostable, Mn2+-dependent 3'-5' exonuclease. The putative polymerase subunit DP2 is not required. The nuclease activity is strongly reduced by single amino acid mutations in the phosphoesterase domain indicating the requirement of this domain for the activity. MjaDP1 acts as a unidirectional, non-processive exonuclease preferring mispaired nucleotides and single-stranded DNA, suggesting that MjaDP1 functions as the proofreading exonuclease of archaeal family D DNA polymerase.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
T. Nuutinen, H. Tossavainen, K. Fredriksson, P. Pirila, P. Permi, H. Pospiech, and J. E. Syvaoja
The solution structure of the amino-terminal domain of human DNA polymerase {varepsilon} subunit B is homologous to C-domains of AAA+ proteins
Nucleic Acids Res., September 1, 2008; 36(15): 5102 - 5110.
[Abstract] [Full Text] [PDF]

Home page
 Microbiol. Mol. Biol. Rev.Home page
E. R. Barry and S. D. Bell
DNA Replication in the Archaea
Microbiol. Mol. Biol. Rev., December 1, 2006; 70(4): 876 - 887.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.