The heterodimeric primase from the euryarchaeon Pyrococcus abyssi: A multifunctional enzyme for initiation and repair?

Type Article
Date 2007-12
Language English
Author(s) Le Breton Magali1, Henneke Ghislaine1, Norais C2, Flament Didier1, Myllykallio H2, Querellou Joel1, Raffin Jean-Paul1
Affiliation(s) 1 : Ifremer, UMR6197, Lab Microbiol Enviornm Extremes, F-29280 Plouzane, France.
2 : Univ Paris 11, Inst Genet & Microbiol, UMR8621, F-91405 Orsay, France.
Source Journal of Molecular Biology (0022-2836) (Elsevier), 2007-12 , Vol. 374 , N. 5 , P. 1172-1185
DOI 10.1016/j.jmb.2007.10.015
WOS© Times Cited 31
Keyword(s) Strand displacement, Gap filling, DNA primase, Archaea, DNA replication
Abstract We report on the characterization of the DNA primase complex of the hyperthermophilic archaeon Pyrococcus abyssi (Pab). The Pab DNA primase complex is composed of the proteins Pabp41 and Pabp46, which show sequence similarities to the p49 and p58 subunits, respectively, of the eukaryotic polymerase alpha-primase complex. Both subunits were expressed, purified, and characterized. The Pabp41 subunit alone had no RNA synthesis activity but could synthesize long (up to 3 kb) DNA strands. Addition of the Pabp46 subunit increased the rate of DNA synthesis but decreased the length of the DNA fragments synthesized and conferred RNA synthesis capability. Moreover, in our experimental conditions, Pab DNA primase had comparable affinities for ribonucleotides and deoxyribonu-cleotides, and its activity was dependent on the presence of Mg2+ and Mn2+. Interestingly, Pab DNA primase also displayed DNA polymerase, gap-filling, and strand-displacement activities. Genetic analyses undertaken in Haloferax volcanii suggested that the eukaryotic-type heterodimeric primase is essential for survival in archaeal cells. Our results are in favor of a multifunctional archaeal primase involved in priming and repair. (c) 2007 Elsevier Ltd. All rights reserved.
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Le Breton Magali, Henneke Ghislaine, Norais C, Flament Didier, Myllykallio H, Querellou Joel, Raffin Jean-Paul (2007). The heterodimeric primase from the euryarchaeon Pyrococcus abyssi: A multifunctional enzyme for initiation and repair? Journal of Molecular Biology, 374(5), 1172-1185. Publisher's official version : https://doi.org/10.1016/j.jmb.2007.10.015 , Open Access version : https://archimer.ifremer.fr/doc/00000/3520/