The nucleotidylylation of herpes simplex virus 1 regulatory protein alpha22 by human casein kinase II.
The products of the alpha genes of herpes simplex virus 1, the infected cells proteins (ICP) 0, 4, 22, and 27 perform regulatory functions, are nucleotidylylated, and share the signaling or recognition sequence (RR(A/T)(P/S)R) that correctly predicted the nucleotidylylation of viral proteins encoded by UL21, UL31, UL49, and UL47 genes expressed later in infection. Extracts from uninfected HeLa cells or casein kinase II purified from sea star nucleotidylylated the ICP22 moiety of a glutathione S-transferase-ICP22 (GST22P) fusion protein with [alpha-32P]ATP or [2-3H]ATP. We report that: (i) Purified HeLa cell casein kinase II specifically labeled a glutathione S-transferase fusion protein containing the amino-terminal 151 amino acids of ICP22 with [2-3H]ATP. (ii) Nucleotidylylation of GST-ICP22 by purified enzyme exhibited positive cooperativity (Hill coefficient of 2 and a K' of 3.7 microM) and a Km = 37.7 microM for ATP. (iii) Nucleotidylylation was inhibited by heparin, casein, or ATPalphaS but not by ATPgammaS. (iv) Mutation of the signaling sequence from RRAPRR to LKAPEK abolished nucleotidylylation. We conclude that nucleotidylylation of proteins by casein kinase II requires the presence of the signaling or recognition sequence, involves the cleavage of the phosphodiester bond between the alpha and beta phosphate, and need not be preceded by phosphorylation.
Mitchell
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The Marjorie B. Kovler Viral Oncology Laboratories, The University of Chicago, Chicago, Illinois 60637, USA.
Blaho
J A
JA
McCormick
A L
AL
Roizman
B
B
D000255:Adenosine Triphosphate / Q000378:metabolism*; D047390:Casein Kinase II; D005982:Glutathione Transferase; D006367:HeLa Cells; D018259:Herpesvirus 1, Human / Q000378:metabolism*; D006801:Humans; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010761:Phosphorus Radioisotopes; D017346:Protein Serine-Threonine Kinases / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins
C057294:ICP22 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D009838:Oligodeoxyribonucleotides; D010761:Phosphorus Radioisotopes; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C078424:EUS1 protein, Equine herpesvirus 1; D000255:Adenosine Triphosphate; D005982:Glutathione Transferase; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases