Myristylation and polylysine-mediated activation of the protein kinase domain of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10).
The amino-terminal domain of the large subunit of herpes simplex virus type 2 (HSV-2) ribonucleotide reductase (ICP10) was previously shown to possess protein kinase (PK) activity that localizes to the cytosolic, cytoskeletal, and plasma membrane fractions. Further studies of the PK domain using computer-assisted sequence analysis have identified a single transmembrane segment and fatty acid incorporation findings indicate that ICP10 is myristylated. Myristylation does not depend on a viral enzyme, since myristic acid is incorporated into ICP10 precipitated from cells transfected with an ICP10 expression vector. It is also incorporated into the 57-kDa protein expressed by the amino-terminal PK expression vector. The myristyl moiety is linked through an amide bond. The basic protein polylysine stimulates the kinase activity and alters its divalent cation requirements resulting in 20- to 40-fold stimulation in the presence of 0.1 mM Mn2+. The PK activity is inhibited by antibody to synthetic peptides corresponding to residues 355-369 and 13-26, respectively, located within, and amino-terminal to, the predicted PK catalytic domain.
Chung
T D
TD
Department of Pharmacology and Experimental Therapeutics, University of Maryland School of Medicine, Baltimore 21201.
Wymer
J P
JP
Kulka
M
M
Smith
C C
CC
Aurelian
L
L
D000595:Amino Acid Sequence; D002460:Cell Line; D004789:Enzyme Activation; D066246:ErbB Receptors / Q000235:genetics; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism*; D010957:Plasmids; D011107:Polylysine / Q000494:pharmacology*; D011487:Protein Conformation; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011972:Receptor, Insulin / Q000235:genetics; D011956:Receptors, Cell Surface / Q000235:genetics; D017451:Receptors, Somatomedin; D012264:Ribonucleotide Reductases / Q000235:genetics / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D018139:Simplexvirus / Q000201:enzymology*; D014162:Transfection
D009227:Myristic Acids; D011956:Receptors, Cell Surface; D017451:Receptors, Somatomedin; D019814:Myristic Acid; D011107:Polylysine; D007334:Insulin-Like Growth Factor I; D012264:Ribonucleotide Reductases; D011494:Protein Kinases; D066246:ErbB Receptors; D011972:Receptor, Insulin