Mapping of a self-interaction domain of the cytomegalovirus protein kinase pUL97.
The human cytomegalovirus-encoded protein kinase pUL97 is a determinant of efficient virus replication and fulfils several regulatory functions. In particular, pUL97 interacts with and phosphorylates viral and cellular proteins. Substrate phosphorylation has regulatory consequences on viral replicative stages such as DNA synthesis, transcription and nuclear capsid egress. pUL97, in accordance with related herpesviral protein kinases, possesses strong autophosphorylation activity. Here, we demonstrate that pUL97 shows a pronounced potential to self-interact. Self-interaction of pUL97 is not dependent on its kinase activity, as seen with a catalytically inactive point mutant. The property of self-interaction maps to the amino acid region 231-280 which is separable from the postulated kinase domain. The detection of high-molecular-mass complexes of pUL97 suggests the formation of dimers and oligomers. Importantly, the analysis of pUL97 mutants by in vitro kinase assays demonstrated a correlation between self-interaction and protein kinase activity, i.e. all mutants lacking the ability to self-interact were negative or reduced in their kinase activity. Thus, our findings provide novel insights into the pUL97 structure-activity relationship suggesting an importance of self-interaction for pUL97 functionality.
Schregel
Vera
V
Institute for Clinical and Molecular Virology, University of Erlangen-Nuremberg, Germany.
Auerochs
Sabrina
S
Institute for Clinical and Molecular Virology, University of Erlangen-Nuremberg, Germany.
Jochmann
Ramona
R
Institute for Clinical and Molecular Virology, University of Erlangen-Nuremberg, Germany.
Maurer
Katja
K
Institute for Clinical and Molecular Virology, University of Erlangen-Nuremberg, Germany.
Stamminger
Thomas
T
Institute for Clinical and Molecular Virology, University of Erlangen-Nuremberg, Germany.
Marschall
Manfred
M
Institute for Clinical and Molecular Virology, University of Erlangen-Nuremberg, Germany.
D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D006801:Humans; D047468:Immunoprecipitation; D009154:Mutation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship
D011494:Protein Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); C075365:ganciclovir kinase