Poliovirus-associated protein kinase: destabilization of the virus capsid and stimulation of the phosphorylation reaction by Zn2+.
The previously described poliovirus-associated protein kinase activity phosphorylates viral proteins VP0 and VP2 as well as exogenous proteins in the presence of Mg2+. In this paper, the effect of Zn2+ on the phosphorylation reaction and the stability of the poliovirus capsid has been studied in detail and compared to that of Mg2+. Phosphorylation patterns of viral and other proteins depend on the divalent cation present. In the presence of Zn2+, phosphorylation of capsid proteins VP2 and VP4 is significantly higher while phosphorylation of VP0 and exogenous phosphate acceptor proteins is not detected. Our results indicate the activation of more than one virus-associated protein kinase by Zn2+. The ion-dependent behavior of the enzyme activities is observed independently of whether the virus was obtained from HeLa or green monkey kidney cells. The poliovirus capsid is destabilized by Zn2+. The destabilization leads to a substantially increased permeability of virus particles to ethidium bromide and RNase, concomitant with decreased infectivity of the sample. This alteration of the poliovirus capsid structure is a prerequisite for effective phosphorylation of viral capsid proteins. The increased level of phosphorylation of viral capsid proteins results in further destabilization of the viral capsid. As a result of the conformational changes, poliovirus-associated protein kinase activities dissociate from the virus particle. High-performance liquid chromatography-purified viral protein VP2 is phosphorylated by the released enzymes on serine, threonine, and tyrosine in the presence of Zn2+. We suggest that the destabilizing effect of phosphorylation on the viral capsid plays a role in uncoating of poliovirus.
Ratka
M
M
Department of Molecular Biology, University of Hamburg, Federal Republic of Germany.
Lackmann
M
M
Ueckermann
C
C
Karlins
U
U
Koch
G
G
D000596:Amino Acids / Q000032:analysis; D002213:Capsid / Q000378:metabolism*; D006801:Humans; D008274:Magnesium / Q000494:pharmacology; D010766:Phosphorylation; D017955:Poliovirus / Q000187:drug effects / Q000378:metabolism*; D011494:Protein Kinases / Q000302:isolation & purification / Q000502:physiology*; D015032:Zinc / Q000494:pharmacology*
D000596:Amino Acids; D011494:Protein Kinases; D008274:Magnesium; D015032:Zinc