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Molecular and Cellular Biology, January 2002, p. 505-516, Vol. 22, No. 2
0270-7306/01/$04.00+0     DOI: 10.1128/MCB.22.2.505-516.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of a Multifunctional Domain in Autonomously Replicating Sequence-Binding Factor 1 Required for Transcriptional Activation, DNA Replication, and Gene Silencing

Tsuyoshi Miyake, Christian M. Loch, and Rong Li*

Department of Biochemistry and Molecular Genetics, School of Medicine, University of Virginia, Charlottesville, Virginia 22908-0733

Received 24 August 2001/ Returned for modification 11 October 2001/ Accepted 16 October 2001

Autonomously replicating sequence-binding factor 1 (ABF1) is a multifunctional, site-specific DNA binding protein that is essential for cell viability in Saccharomyces cerevisiae. ABF1 plays a direct role in transcriptional activation, stimulation of DNA replication, and gene silencing at the mating-type loci. Here we demonstrate that all three activities of ABF1 are conferred by the C terminus of the protein (amino acids [aa] 604 to 731). Furthermore, a detailed mutational analysis has revealed two important clusters of amino acid residues in the C terminus (C-terminal sequence 1 [CS1], aa 624 to 628; and CS2, aa 639 to 662). While both regions play a pivotal role in supporting cell viability, they make distinct contributions to ABF1 functions in various nuclear processes. CS1 specifically participates in transcriptional silencing and/or repression in a context-dependent manner, whereas CS2 is universally required for all three functions of ABF1. When tethered to specific regions of the genome, a 30-aa fragment that contains CS2 alone is sufficient for activation of transcription and chromosomal replication. In addition, CS2 is responsible for ABF1-mediated chromatin remodeling. Based on these results, we suggest that ABF1 may function as a chromatin-reorganizing factor to increase accessibility of the local chromatin structure, which in turn facilitates the action of additional factors to establish either an active or repressed chromatin state.

* Corresponding author. Mailing address: Department of Biochemistry & Molecular Genetics, School of Medicine, P.O. Box 800733, University of Virginia, Charlottesville, VA 22908-0733. Phone: (804) 243-2727. Fax: (804) 924-5069. E-mail: rl2t{at}virginia.edu.

Molecular and Cellular Biology, January 2002, p. 505-516, Vol. 22, No. 2
0022-538X/01/$04.00+0     DOI: 10.1128/MCB.22.2.505-516.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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