RRC ID 42054
Author Takahashi-Omoe H, Omoe K, Sakaguchi M, Kameoka Y, Matsushita S, Inada T.
Title Analysis of protein expression by mammalian cell lines stably expressing lactate dehydrogenase-elevating virus ORF 5 and ORF 6 proteins.
Journal Comp Immunol Microbiol Infect Dis
Abstract Lactate dehydrogenase-elevating virus (LDV) has a strict species-specificity. Because only a subset of mouse primary macrophages have been identified that can support LDV replication in vitro, the precise molecular mechanism of viral entry and replication remains unclear. To analyze the LDV envelope proteins, which probably mediate viral attachment to the host cell, we developed a mammalian system for stable co-expression of LDV open reading frame (ORF) 5- and ORF 6-encoded proteins (ORF 5 and ORF 6 proteins), which correspond to envelope VP-3 and M/VP-2, respectively, and compared these expressed proteins to the native ones. Western blotting analysis combined with N-glycanase digestion revealed that ORF 5 and ORF 6 proteins were similar in size to native VP-3 and M/VP-2, and that ORF 5 protein was N-glycosylated, like the native VP-3. Immunofluorescence microscopy revealed that both ORF 5 and ORF 6 proteins were distributed throughout the cytoplasm and were colocalized in most cells. Moreover, ORF 5 protein was localized both in the perinuclear region and the Golgi complex and transported to the cell surface. This mammalian expression system in which the exogenously expressed proteins closely resemble the native proteins will provide the experimental basis for further studies of the interactions between LDV envelope proteins and host cells.
Volume 27(2)
Pages 81-92
Published 2004-3-1
DOI 10.1016/S0147-9571(03)00053-5
PII S0147-9571(03)00053-5
PMID 14690718
PMC PMC7124688
MeSH Animals Blotting, Western COS Cells Chlorocebus aethiops Gene Expression Regulation, Viral / physiology* Lactate dehydrogenase-elevating virus / genetics Lactate dehydrogenase-elevating virus / metabolism* Membrane Glycoproteins Microscopy, Fluorescence Open Reading Frames Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / metabolism Transfection Viral Envelope Proteins / biosynthesis* Viral Envelope Proteins / genetics
IF 1.573
Times Cited 1
WOS Category VETERINARY SCIENCES IMMUNOLOGY MICROBIOLOGY
Resource
Human and Animal Cells COS-7(RCB0539)