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Originally published In Press as doi:10.1074/jbc.M101537200 on April 6, 2001

J. Biol. Chem., Vol. 276, Issue 26, 23440-23449, June 29, 2001
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Novel Sp Family-like Transcription Factors Are Present in Adult Insect Cells and Are Involved in Transcription from the Polyhedrin Gene Initiator Promoter*

Aruna RamachandranDagger §, Anjali Jain§||, Puneeta Arora**, Murali Dharan BashyamDagger §, Udayan ChatterjeeDagger Dagger Dagger , Sudip Ghosh§, Veena K. Parnaik**, and Seyed E. HasnainDagger §§§

From the Dagger  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

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.

* 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.

These authors contributed equally to this work.

|| Present address: Howard Hughes Medical Inst., UCLA School of Medicine, Los Angeles, CA 90024-1662.

Dagger Dagger 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.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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