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J. Biol. Chem., Vol. 279, Issue 32, 33673-33683, August 6, 2004

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A Protein Sequestering System Reveals Control of Cellular Programs by the Transcriptional Coactivator HCF-1^*

Bharat Khurana and Thomas M. Kristie

From the Laboratory of Viral Diseases, National Institutes of Health, Bethesda, Maryland 20892

The mammalian transcriptional coactivator HCF-1 is a critical component of the multiprotein herpes simplex virus immediate early gene enhancer core complex. The protein has also been implicated in basic cellular processes such as cell-cycle progression, transcriptional coactivation, and mRNA processing. Functions have been attributed to HCF-1 primarily from analyses of protein-protein interactions and from the cell-cycle-arrested phenotype of an HCF-1 temperature-sensitive mutant. However, neither the mechanisms involved nor specific cellular transcriptional targets have been identified. As the protein is essential for cell viability and proliferation, a genetic system was developed to specifically sequester the nuclear factor in the cell cytoplasm in a regulated manner. This approach exhibits no significant cell toxicity yet clearly demonstrates the requirement of available nuclear HCF-1 for herpes simplex virus immediate early gene expression during productive infection. Additionally, cellular transcriptional events were identified that contribute to understanding the functions ascribed to the protein and implicate the protein in events that impact the regulation of critical cellular processes.

Received for publication, February 4, 2004 , and in revised form, June 7, 2004.

^* This work was supported by the Laboratory of Viral Diseases, NIAID, National Institutes of Health (to T. M. K.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

This article was selected as a Paper of the Week.

To whom correspondence should be addressed: NIH, Bldg. 4, Rm. 131, 4 Center Dr., Bethesda, MD 20892. Tel.: 301-496-3854; Fax: 301-480-1560; E-mail: thomas_Kristie{at}nih.gov.

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