Origin of expression of the herpes simplex virus type 1 protein U(S)1.5.
ICP22, an immediate-early protein of herpes simplex virus type 1 (HSV-1), is required for viral replication in nonpermissive cell types and for expression of a class of late viral proteins which includes glycoprotein C. An understanding of the mechanism of ICP22 function has been complicated by the coexpression of the full-length protein with an in-frame, C-terminus-specific protein, U(S)1.5. In this report, we confirm that the U(S)1.5 protein is a bona fide translation product since it is detected during infections with three laboratory strains and two low-passage clinical isolates of HSV-1. To clarify the expression patterns of the ICP22 and U(S)1.5 proteins, we examined their synthesis from plasmids in transient expression assays. Because previous studies had identified two different U(S)1.5 translational start sites, we attempted to determine which is correct by studying the effects of a series of deletion, nonsense, and methionine substitutions on U(S)1.5 expression. First, amino acids 90 to 420 encoded by the ICP22 open reading frame (ORF) migrated at the mobility of U(S)1.5 in sodium dodecyl sulfate-polyacrylamide gels. Second, introduction of a stop codon downstream of M90 ablated expression of both ICP22 and U(S)1.5. Finally, mutation of M90 to alanine (M90A) allowed expression of full-length ICP22 while dramatically reducing expression of U(S)1.5. Levels of U(S)1.5 but not ICP22 protein expression were also reduced in cells infected with an M90A mutant virus. Thus, we conclude that expression of IC22 and that of U(S)1.5 can occur independently of each other and that U(S)1.5 translation initiates at M90 of the ICP22 ORF.
Bowman
J Jason
JJ
Department of Medicine and Microbiology, Harvard Medical School at the Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215, USA.
Schaffer
Priscilla A
PA
D018259:Herpesvirus 1, Human / Q000502:physiology*; D017874:Immediate-Early Proteins / Q000235:genetics*; D009154:Mutation; D016366:Open Reading Frames / Q000235:genetics; D014176:Protein Biosynthesis; D014764:Viral Proteins / Q000096:biosynthesis*; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics
C057294:ICP22 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins