Nucleic Acids Research, Vol 27, Issue 22 4444-4450, Copyright © 1999 by Oxford University Press
G Desogus, S Onesti, P Brick, M Rossi and FM Pisani
We report the identification and characterisation of a DNA primase from the
thermophilic methanogenic archaeon Methanococcus jannaschii (Mjpri). The
analysis of the complete genome sequence of this organism has identified an
open reading frame coding for a protein with sequence similarity to the
small subunit of the eukaryotic DNA primase (the p50 subunit of the
polymerase alpha-primase complex). This protein has been overexpressed in
Escherichia coli and purified to near homogeneity. Recombinant Mjpri is
able to synthesise oligoribonucleotides on various pyrimidine
single-stranded DNA templates [poly(dT) and poly(dC)]. This activity
requires divalent cations such Mg(2+), Mn(2+)or Zn(2+), and is additionally
stimulated by the monovalent cation K(+). A multiple sequence alignment has
revealed that most of the regions that are conserved in eukaryotic p50
subunits are also present in the archaeal primases, including the conserved
negatively charged residues, which have been shown to be essential for
catalysis in the mouse primase. Of the four cysteine residues that have
been postulated to make up a putative Zn-binding motif, two are not present
in the archaeal homologue. This is the first report on the biochemical
characterisation of an archaeal DNA primase.
ARTICLES
Identification and characterization of a DNA primase from the hyperthermophilic archaeon Methanococcus jannaschii
Imperial College of Science, Technology, and Medicine, Biophysics Section, Blackett Laboratory, London SW7 2BZ, UK.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  L. Yakovleva and S. Shuman Nucleotide Misincorporation, 3'-Mismatch Extension, and Responses to Abasic Sites and DNA Adducts by the Polymerase Component of Bacterial DNA Ligase D J. Biol. Chem., September 1, 2006; 281(35): 25026 - 25040. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. De Falco, A. Fusco, M. De Felice, M. Rossi, and F. M. Pisani The DNA primase of Sulfolobus solfataricus is activated by substrates containing a thymine-rich bubble and has a 3'-terminal nucleotidyl-transferase activity Nucleic Acids Res., September 30, 2004; 32(17): 5223 - 5230. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Seybert, D. J. Scott, S. Scaife, M. R. Singleton, and D. B. Wigley Biochemical characterisation of the clamp/clamp loader proteins from the euryarchaeon Archaeoglobus fulgidus Nucleic Acids Res., October 15, 2002; 30(20): 4329 - 4338. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Liu, K. Komori, S. Ishino, A. A. Bocquier, I. K. O. Cann, D. Kohda, and Y. Ishino The Archaeal DNA Primase. BIOCHEMICAL CHARACTERIZATION OF THE p41-p46 COMPLEX FROM PYROCOCCUS FURIOSUS J. Biol. Chem., November 21, 2001; 276(48): 45484 - 45490. [Abstract] [Full Text] [PDF]
|
|