Novel Sp family-like transcription factors are present in adult insect cells and are involved in transcription from the polyhedrin gene initiator promoter.
We earlier documented the involvement of a cellular factor, polyhedrin (polh) promoter-binding protein, in transcription from the Autographa californica nuclear polyhedrosis virus polh gene promoter. Sequences upstream of the polh promoter were found to influence polh promoter-driven transcription. Analysis of one such region, which could partially compensate for the mutated polh promoter and also activate transcription from the wild-type promoter, revealed a sequence (AcSp) containing a CACCC motif and a loose GC box resembling the binding motifs of the transcription factor Sp1. AcSp and the consensus Sp1 sequence (cSp) specifically bound factor(s) in HeLa and Spodoptera frugiperda (Sf9) insect cell nuclear extracts to generate identical binding patterns, indicating the similar nature of the factor(s) interacting with these sequences. The AcSp and cSp oligonucleotides enhanced in vivo expression of a polh promoter-driven luciferase gene. In vivo mopping of these factor(s) significantly reduced transcription from the polh promoter. Recombinant viruses carrying deletions in the upstream AcSp sequence confirmed the requirement of these factor(s) in polh promoter-driven transcription in the viral context. We demonstrate for the first time DNA-protein interactions involving novel members of the Sp family of proteins in adult insect cells and their involvement in transcription from the polh promoter.
Ramachandran
A
A
Eukaryotic Gene Expression Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110067, India.
Jain
A
A
Arora
P
P
Bashyam
M D
MD
Chatterjee
U
U
Ghosh
S
S
Parnaik
V K
VK
Hasnain
S E
SE
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D002457:Cell Extracts / Q000032:analysis; D002467:Cell Nucleus / Q000378:metabolism; D016384:Consensus Sequence; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D017930:Genes, Reporter; D005814:Genes, Viral; D006367:HeLa Cells; D006801:Humans; D019476:Insect Proteins; D009687:Nuclear Proteins / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D000077183:Occlusion Body Matrix Proteins; D011401:Promoter Regions, Genetic; D016329:Sp1 Transcription Factor / Q000502:physiology*; D018411:Spodoptera / Q000378:metabolism / Q000821:virology*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics*; D015678:Viral Structural Proteins; D015032:Zinc / Q000378:metabolism
D002457:Cell Extracts; D004268:DNA-Binding Proteins; D019476:Insect Proteins; D009687:Nuclear Proteins; D000077183:Occlusion Body Matrix Proteins; D016329:Sp1 Transcription Factor; D014764:Viral Proteins; D015678:Viral Structural Proteins; C085605:polyhedrin promoter-binding protein, insect; C009829:polyhedrin protein, Nucleopolyhedrovirus; D004247:DNA; D015032:Zinc