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Virus Genes. 2015 Oct;51(2):171-81. doi: 10.1007/s11262-015-1233-6. Epub 2015 Aug 21.

Production and characterisation of Epstein-Barr virus helicase-primase complex and its accessory protein BBLF2/3.

Author information

1
Univ. Grenoble Alpes, UVHCI, 38000, Grenoble, France.
2
CNRS, UVHCI, 38000, Grenoble, France.
3
European Synchrotron Radiation Facility (ESRF), 38000, Grenoble, France.
4
EMBL Grenoble Outstation, UVHCI, 38000, Grenoble, France.
5
Laboratoire de Virologie, Centre Hospitalo-Universitaire de Grenoble, B.P. 217, 38043, Grenoble Cedex 9, France.
6
Univ. Grenoble Alpes, UVHCI, 38000, Grenoble, France. wim.burmeister@ujf-grenoble.fr.
7
CNRS, UVHCI, 38000, Grenoble, France. wim.burmeister@ujf-grenoble.fr.
8
Univ. Grenoble Alpes, UVHCI, 38000, Grenoble, France. hutin@uvhci.fr.
9
CNRS, UVHCI, 38000, Grenoble, France. hutin@uvhci.fr.

Abstract

The helicase-primase complex is part of the lytic DNA replication machinery of herpesviruses, but up to now, almost nothing is known about its structure. For Epstein-Barr virus it consists in the helicase BBLF4, the primase BSLF1 and the accessory protein BBLF2/3. The accessory protein shows only weak sequence homology within the herpesvirus family but may be related to an inactive B-family polymerase. BSLF1 belongs to the archaeo-eukaryotic primase family, whereas the helicase BBLF4 has been related either to Dda helicases of caudovirales or to Pif1 helicases. We produced the helicase-primase complex in insect cells using a baculovirus coding for all three proteins simultaneously. The soluble monomeric helicase-primase complex containing the three proteins with 1:1:1 stoichiometry showed ATPase activity, which is strongly stimulated in the presence of ssDNA oligomers. Furthermore, we expressed BBLF2/3 as soluble monomeric protein and performed small-angle X-ray scattering experiments which yielded an envelope whose shape is compatible with B-family polymerases.

KEYWORDS:

DNA helicase; DNA replication; EBV; Epstein–Barr virus; Gammaherpesvirus; Primase

PMID:
26292944
DOI:
10.1007/s11262-015-1233-6
[Indexed for MEDLINE]

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