Promoter analysis of the Chilo iridescent virus DNA polymerase and major capsid protein genes.
The DNA polymerase (DNApol) and major capsid protein (MCP) genes were used as models to study promoter activity in Chilo iridescent virus (CIV). Infection of Bombyx mori SPC-BM-36 cells in the presence of inhibitors of DNA or protein synthesis showed that DNApol, as well as helicase, is an immediate-early gene and confirmed that the major capsid protein (MCP) is a late gene. Transcription of DNApol initiated 35 nt upstream and that of MCP 14 nt upstream of the translational start site. In a luciferase reporter gene assay both promoters were active only when cells were infected with CIV. For DNApol sequences between position -27 and -6, relative to the transcriptional start site, were essential for promoter activity. Furthermore, mutation of a G within the sequence TTGTTTT located just upstream of the DNApol transcription initiation site reduced the promoter activity by 25%. Sequences crucial for MCP promoter activity are located between positions -53 and -29.
Nalçacioğlu
Remziye
R
Laboratory of Virology, Wageningen University, Binnenhaven 11, 6709 PD, Wageningen, The Netherlands.
Marks
Hendrik
H
Vlak
Just M
JM
Demirbaĝ
Zihni
Z
van Oers
Monique M
MM
D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000821:virology; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism; D017994:Iridovirus / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
D036022:Capsid Proteins; D017874:Immediate-Early Proteins; D008156:Luciferases; D004259:DNA-Directed DNA Polymerase; D004265:DNA Helicases