Conformation-defective herpes simplex virus 1 glycoprotein B activates the promoter of the grp94 gene that codes for the 94-kD stress protein in the endoplasmic reticulum.
GRP94 is a major glycoprotein in the endoplasmic reticulum with calcium-binding properties. Recently, GRP94 has been shown to bind to unassembled forms of multimeric proteins and peptides. We report here that GRP94 forms a stable association with the mutated form of the herpes simplex type virus 1 (HSV-1) glycoprotein B, but not with the fully processed viral protein. Both the glycosylated and unglycosylated forms of GRP94 are capable of complexing with the mutated, conformation-defective viral glycoprotein. Cotransfection of expression vectors for gB and grp94 promoter fusion genes revealed that the grp94 promoter is strongly activated by the mutant form of gB. Analysis of the grp94 promoter mutants showed that two regions in the promoter, a highly conserved element referred to as grp core and the CCAAT element most proximal to the TATA element (C1), mediate the induction of grp94 by malfolded protein. We further determined that the grp94 core and C1 element bind to common as well distinct nuclear factors from grp78, a commonly coregulated gene. Through UV cross-linking, site competition, and immunocross-reactivity, we identified that the heteromeric CCAAT-binding protein (CBF) is one component of the grp94 C1 complex.
Ramakrishnan
M
M
Department of Biochemistry and Molecular Biology, University of Southern California School of Medicine, Los Angeles 90033, USA.
Tugizov
S
S
Pereira
L
L
Lee
A S
AS
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004721:Endoplasmic Reticulum / Q000737:chemistry; D005786:Gene Expression Regulation; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics*; D018832:Molecular Chaperones / Q000737:chemistry; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D011401:Promoter Regions, Genetic / Q000187:drug effects; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D014162:Transfection; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000494:pharmacology
D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D018832:Molecular Chaperones; D014759:Viral Envelope Proteins; C018551:glucose-regulated proteins; C040495:glycoprotein B, Simplexvirus