Expression and localization of LEF-11 in Autographa californica nucleopolyhedrovirus-infected Sf9 cells.
The Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) lef-11 gene was found previously to be necessary to support optimal levels of transient expression from an AcMNPV late promoter. The lef-11 gene is unusual in that it overlaps both upstream (orf38) and downstream (pp31) genes. In this study, the expression and cellular localization of LEF-11 were examined. The lef-11 transcripts were detected from 4 to 36 h post-infection (p.i.). The 1.5 kb lef-11 mRNA initiates 196 nt upstream of the lef-11 translation initiation codon, within the upstream orf38 gene. This relatively long 5' upstream region encodes a potential small upstream open reading frame (ORF) of 58 amino acids that overlaps the lef-11 ORF. The 3' end of the lef-11 mRNA was mapped as co-terminal with mRNAs from the downstream pp31 gene. Using affinity purified anti-LEF-11 antibodies, levels of LEF-11 expression were found to be maximal between approximately 8 and 24 h p.i., although LEF-11 could be detected as late as 72 h p.i. Using immunofluorescence microscopy, it was determined that LEF-11 localized to dense regions of infected cell nuclei, consistent with its role as a possible late transcription factor.
Lin
Guangyun
G
Boyce Thompson Institute at Cornell University, Tower Road, Ithaca, NY 14853-1801, USA1.
Slack
Jeffrey M
JM
Boyce Thompson Institute at Cornell University, Tower Road, Ithaca, NY 14853-1801, USA1.
Blissard
Gary W
GW
Boyce Thompson Institute at Cornell University, Tower Road, Ithaca, NY 14853-1801, USA1.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016366:Open Reading Frames; D012333:RNA, Messenger / Q000032:analysis; D018411:Spodoptera; D014764:Viral Proteins / Q000032:analysis* / Q000096:biosynthesis
D012333:RNA, Messenger; D014764:Viral Proteins