The host factor polyhedrin promoter binding protein (PPBP) is involved in transcription from the baculovirus polyhedrin gene promoter

Sudip Ghosh, Anjali Jain, Bipasha Mukherjee, Saman Habib, Seyed E. Hasnain

Research output: Contribution to journalArticle

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Abstract

Hypertranscription and temporal expression from the Autographa californica nuclear polyhedrosis (AcNPV) baculovirus polyhedrin promoter involves an α-amanitin-resistant RNA polymerase and requires a trans-acting viral factor(s). We previously reported that a 30-kDa host factor, polyhedrin promoter binding protein (PPBP), binds with unusual affinity, specificity, and stability to the transcriptionally important motif AATAA ATAAGTATT within the polyhedrin (polh) initiator promoter and also displays coding strand- specific single-stranded DNA (ssDNA)-binding activity (S. Burma, B. Mukherjee, A. Jain, S. Habib, and S. E. Hasnain, J. Biol. Chem. 269:2750- 2757, 1994; B. Mukherjee, S. Burma, and S. E. Hasnain, J. Biol. Chem. 270:4405-4411, 1995). We now present evidence which indicates that an additional factor(s) is involved in stabilizing PPBP-duplex promoter and PPBP-ssDNA interactions. TBP (TATA box binding protein) present in Spodoptera frugiperda (Sf9) cells is characteristically distinct from PPBP and does not interact directly with the polh promoter. RePlacement of PPBP cognate sequences within the polh promoter with random nucleotides abolished PPBP binding in vitro and also failed to express the luciferase reporter gene in vivo. Phosphocellulose fractions of total nuclear extract from virus-infected cells which support in vitro transcription from the polh promoter contain PPBP activity. When PPBP was sequestered by the presence of oligonucleotides containing PPBP cognate sequence motifs, in vitro transcription of a C-free reporter cassette was affected but was restored by the exogenous addition of nuclear extract containing PPBP. When PPBP was mopped out in vivo by a plasmid carrying PPBP cognate sequence present in trans, polh promoter- driven expression of the luciferase reporter was abolished, demonstrating that binding of PPBP to the polh promoter is essential for transcription.

Original languageEnglish (US)
Pages (from-to)7484-7493
Number of pages10
JournalJournal of Virology
Volume72
Issue number9
StatePublished - Sep 1998

Fingerprint

Baculoviridae
binding proteins
Carrier Proteins
transcription (genetics)
promoter regions
Genes
genes
Luciferases
Amanitins
single-stranded DNA
TATA-Box Binding Protein
amino acid sequences
Sf9 Cells
Spodoptera
luciferase
Trans-Activators
Single-Stranded DNA
DNA-Binding Proteins
DNA-Directed RNA Polymerases
Reporter Genes

ASJC Scopus subject areas

  • Immunology

Cite this

The host factor polyhedrin promoter binding protein (PPBP) is involved in transcription from the baculovirus polyhedrin gene promoter. / Ghosh, Sudip; Jain, Anjali; Mukherjee, Bipasha; Habib, Saman; Hasnain, Seyed E.

In: Journal of Virology, Vol. 72, No. 9, 09.1998, p. 7484-7493.

Research output: Contribution to journalArticle

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