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J. Biol. Chem., Vol. 277, Issue 9, 6943-6948, March 1, 2002
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From the Institut für Klinische und Molekulare Virologie,
D-91054 Erlangen, Germany
Herpesviral DNA packaging is a complex process
resulting in unit-length genomes packed into preformed procapsids. This
process is believed to be mediated by two packaging proteins, the
terminase subunits. In the case of double-stranded DNA
bacteriophages, the translocation of DNA was shown to be an
energy-dependent process associated with an ATPase activity
of the large terminase subunit. In the case of human cytomegalovirus it
was not known which protein has the ability to hydrolyze ATP. In this
study we expressed human cytomegalovirus terminase subunits, pUL89 and
the carboxyl-terminal half of pUL56, as GST fusion proteins and
purified these by affinity chromatography. ATPase assays demonstrated
that the enzymatic activity is exclusively associated with pUL56. The
characterization of the ATP hydrolysis showed that the enzymatic
reaction is a fast process, whereas the spontaneous ATP decay followed
slow kinetics. Interestingly, although pUL89 did not show any ATPase activity, it was capable of enhancing the UL56-associated ATP hydrolysis. Furthermore, a specific association of in vitro
translated pUL89 with the carboxyl-terminal half of GST-UL56C was
detected. This interaction was confirmed by co-immunoprecipitations of
infected cells. Our results clearly demonstrated that (i) both
terminase subunits interact with each other and (ii) the subunit pUL56
has an ATPase activity.
ATPase Activity of the Terminase Subunit pUL56 of Human
Cytomegalovirus*
*
This study was supported by the Johannes and Frieda
Marohn Foundation.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Recipient of a Habilitationstipendium from the Deutsche
Forschungsgemeinschaft. To whom correspondence should be addressed: Institut für Klinische und Molekulare Virologie, Schlossgarten 4, 91054 Erlangen, Germany. Tel.: 49-9131-8522104; Fax: 49-9131-8526493; E-mail: eebogner@viro.med.uni-erlangen.de.
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