Site-directed mutation in a conserved kinase domain of human cytomegalovirus-pp65 with preservation of cytotoxic T lymphocyte targeting.
The major target of human cytomegalovirus (CMV)-specific cytotoxic T lymphocytes (CTL) is the tegument protein CMVpp65. However, this protein has protein kinase (PK) activity, and the unknown effects on cell replication of an exogenous PK in healthy cells could limit the use of CMVpp65 as a vaccine, especially in children. In this report we show that a point mutation converting lysine to asparagine at the invariant lysine (K436), an essential site for phosphotransfer, abolishes the threonine kinase activity. The mutant CMVpp65 maintains its immunologic target characteristics, including antibody and CTL reactivity. This kinase-deficient CMVpp65 is a candidate for evaluation in future CMV vaccine development.
Yao
Z Q
ZQ
Department of Virology, Beckman Research Institute of the City of Hope, 1500 East Duarte Road, Duarte, CA 91010, USA.
Gallez-Hawkins
G
G
Lomeli
N A
NA
Li
X
X
Molinder
K M
KM
Diamond
D J
DJ
Zaia
J A
JA
D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D002364:Caseins / Q000378:metabolism; D020134:Catalytic Domain; D002460:Cell Line; D002857:Chromium / Q000378:metabolism; D017124:Conserved Sequence / Q000235:genetics*; D003587:Cytomegalovirus / Q000201:enzymology / Q000235:genetics / Q000276:immunology*; D022482:Cytomegalovirus Vaccines / Q000235:genetics / Q000276:immunology*; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D015789:HLA-A2 Antigen / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D010766:Phosphorylation; D010769:Phosphothreonine / Q000378:metabolism; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
D000956:Antigens, Viral; D002364:Caseins; D022482:Cytomegalovirus Vaccines; D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; D010750:Phosphoproteins; D014614:Vaccines, Synthetic; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D002857:Chromium; D010769:Phosphothreonine; D011494:Protein Kinases