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Mol Cell Biol, March 1998, p. 1296-1302, Vol. 18, No. 3
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Activation of Chromosomal DNA Replication in Saccharomyces cerevisiae by Acidic Transcriptional Activation Domains

Rong Li,1,2,* David S. Yu,1 Masafumi Tanaka,2,dagger Liyi Zheng,1 Shelley L. Berger,3 and Bruce Stillman2

Department of Biochemistry and Molecular Genetics, Health Sciences Center, University of Virginia, Charlottesville, Virginia 229081; Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 117242; and The Wistar Institute, Philadelphia, Pennsylvania 191043

Received 9 September 1997/Returned for modification 17 November 1997/Accepted 9 December 1997

A large body of evidence from viral systems has established that transcription factors play an important and direct role in activating viral DNA replication. Among the transcriptional activation domains that can stimulate viral DNA replication are acidic domains such as those derived from herpes simplex virus VP16 and the tumor suppressor p53. Here we show that acidic activation domains can also activate a cellular origin of replication in a chromosomal context. When tethered to the yeast ARS1 (autonomously replicating sequence 1) origin of replication, both VP16 and p53 activation domains can enhance origin function. In addition, the C-terminal acidic region of the yeast transcription factor ABF1, which normally activates the ARS1 origin, is sufficient for activating ARS1 function when tethered to the origin. Mutations at residues Trp-53 and Phe-54 of a 20-residue (41 to 60) activation region of p53 abolish the activation of both replication and transcription, suggesting that the same structural determinants may be employed to activate both processes in yeast. Furthermore, using a two-dimensional gel electrophoresis method, we demonstrate that the GAL4-p53 chimeric activator can activate initiation of chromosomal replication from an origin inserted at the native ARS1 locus. These findings strongly suggest functional conservation of the mechanisms used by the acidic activation domains to activate viral DNA replication in mammalian cells and chromosomal replication in yeast.

* Corresponding author. Mailing address: Department of Biochemistry, Health Sciences Center, University of Virginia, Charlottesville, VA 22908. Phone: (804) 243-2727. Fax: (804) 924-5069. E-mail: rl2t{at}virginia.edu.

dagger Present address: Molecular Medicine Research Center, The Institute of Medical Science, Tokai University, Bohseida, Isehara, Kanagawa 259-11, Japan.




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