ICP27 phosphorylation site mutants are defective in herpes simplex virus 1 replication and gene expression.
Herpes simplex virus 1 (HSV-1) protein ICP27 is a multifunctional regulatory protein that is posttranslationally modified by phosphorylation during viral infection. ICP27 has been shown to be phosphorylated on three serine residues, specifically serine residues 16 and 18, which are within casein kinase 2 (CK2) sites, and serine residue 114, which is within a protein kinase A (PKA) site. Phosphorylation is an important regulatory mechanism that is reversible and controls many signaling pathways, protein-protein interactions, and protein subcellular localization. To determine the role of phosphorylation in modulating the activities of ICP27, we constructed phosphorylation site mutations at each of the three serine residues. Single, double, and triple viral mutants were created in which alanine or glutamic acid was substituted for serines 16, 18, and 114. ICP27 phosphorylation site mutants were defective in viral replication and viral gene expression. Notably, ICP4-containing replication compartment formation was severely compromised, with the appearance of small ring-like structures that persisted even at late times after infection. Neither the colocalization of ICP27 with RNA polymerase II nor the formation of Hsc70 nuclear foci was observed during infection with the phosphorylation site mutants, both of which occur during wild-type HSV-1 infection. These data indicate that several key events in which ICP27 plays a role are curtailed during infection with ICP27 phosphorylation site mutants.
Rojas
Santos
S
Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Irvine, California 92697-4025, USA.
Corbin-Lickfett
Kara A
KA
Escudero-Paunetto
Laurimar
L
Sandri-Goldin
Rozanne M
RM
D000818:Animals; D002460:Cell Line; D015967:Gene Expression Regulation, Viral; D050883:HSC70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics* / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D012694:Serine / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics*
D050883:HSC70 Heat-Shock Proteins; C057120:ICP27 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D012694:Serine; D012319:RNA Polymerase II