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J. Biol. Chem., Vol. 276, Issue 26, 23440-23449, June 29, 2001
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From the 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.
Novel Sp Family-like Transcription Factors Are Present in Adult
Insect Cells and Are Involved in Transcription from the Polyhedrin Gene
Initiator Promoter*
§¶,
,§,,§§§
Eukaryotic Gene Expression Laboratory,
National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi
110067, India, the § Laboratory of Molecular and Cellular
Biology, Centre for DNA Fingerprinting and Diagnostics, ECIL
Road, Nacharam, Hyderabad 500076, India, and the ** Centre for Cellular
and Molecular Biology, Uppal Road, Hyderabad 500007, India
*
This work was supported by Department of Science and
Technology Grant SP/SO/D-45/95 (to S. E. H.) and by core
support from the Department of Biotechnology, Government of India to
National Institute of Immunology and Centre for DNA Fingerprinting and Diagnostics.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Present address: Howard Hughes Medical Inst., UCLA School of
Medicine, Los Angeles, CA 90024-1662.
Present address: Dept. of Molecular Biology, Scripps Research
Inst., 10550 N. Torrey Pines Rd., La Jolla, CA 92037.
§§
To whom correspondence should be addressed. Tel.:
91-40-7155604; Fax: 91-40-7155610; E-mail:
ehtesham@www.cdfd.org.in.
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