Human cytomegalovirus carries serine/threonine protein phosphatases PP1 and a host-cell derived PP2A.
Human cytomegalovirus (CMV), a herpesvirus, is an important cause of morbidity and mortality in immunocompromised patients. When studying hyper-immediate-early events after contact between CMV virions and the cell membrane, we observed a hypophosphorylation of cellular proteins within 10 min. This can be explained in part by our finding that purified CMV contains serine/threonine protein phosphatase activities. Biochemical analyses indicate that this protein phosphatase activity has all characteristics of type 1 and 2A protein phosphatases (PP1 and PP2A). Specifically, PP1 accounts for approximately 30% and PP2A accounts for the remaining 70% of the phosphorylase phosphatase activity found. CMV produced in astrocytoma cells stably expressing an amino-terminally tagged PP2A catalytic subunit contained tagged enzyme, thus demonstrating the cellular origin of CMV-associated PP2A. PP2A is specifically found inside the virus, associated with the nucleocapsid fraction. Western blot (immunoblot) analysis of purified virus revealed the presence of the catalytic subunits of PP2A and PP1. Furthermore, the catalytic subunit of PP2A appears to be complexed to the regulatory subunits PR65 and PR55, which is also the most abundant configuration of this enzyme found in the host cells. Incubation of virus with okadaic acid before contact of CMV with cells prevented hypophosphorylation of cellular proteins, thus demonstrating the role of CMV-associated phosphatases in this phenomenon. CMV can thus transport an active enzyme from one cell to another.
Michelson
S
S
Unité d'Immunologie Virale, Institut Pasteur, Paris, France.
Turowski
P
P
Picard
L
L
Goris
J
J
Landini
M P
MP
Topilko
A
A
Hemmings
B
B
Bessia
C
C
Garcia
A
A
Virelizier
J L
JL
D000595:Amino Acid Sequence; D002460:Cell Line; D003587:Cytomegalovirus / Q000201:enzymology* / Q000648:ultrastructure; D006801:Humans; D008969:Molecular Sequence Data; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation; D014407:Tumor Cells, Cultured; D014771:Virion / Q000378:metabolism
D010749:Phosphoprotein Phosphatases