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Role of multifunctional autonomously replicating sequence binding factor 1 in the initiation of DNA replication and transcriptional control in Saccharomyces cerevisiae

Rhode, Peter R. and Elasser, Suzanne and Campebell, Judith L. (1992) Role of multifunctional autonomously replicating sequence binding factor 1 in the initiation of DNA replication and transcriptional control in Saccharomyces cerevisiae. Molecular and Cellular Biology, 12 (3). pp. 1064-1077. ISSN 0270-7306. http://resolver.caltech.edu/CaltechAUTHORS:RHOmcb92

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Abstract

Autonomously replicating sequence (ARS) binding factor 1 (ABF1) is an abundant DNA-binding protein that specifically recognizes the motif RTCRYN5ACG at many sites in the yeast genome, including promoter elements, mating-type silencers, and ARSs. Mutational analysis of these sites suggests that ABF1 is involved in constitutive and carbon source-regulated transcriptional activation, transcriptional silencing, and ARS activity. To better assess the role of ABF1 in DNA replication and transcriptional control, temperature-sensitive lethal mutations in the ABF1 gene were isolated. Several of the abf1(Ts) strains show rapid growth arrest at the nonpermissive temperature. At the semipermissive temperature, these strains show an ARS-specific defect in the mitotic stability of ARS-CEN plasmids, such that the abf1 mutants show defects in ARS function identical to those of mutants bearing the mutations in the cis-acting ABF1 binding sites analyzed previously by numerous investigators. Flow cytometric analysis and in vivo DNA labeling experiments on an alpha-factor synchronized abf1(Ts) strain showed that at the nonpermissive temperature, these cells fail to progress efficiently from G1 through S phase and synthesize DNA at 25% of the level seen in the isogenic ABF1 strain. RNA synthesis is also reduced in the abf1(Ts) strains. In addition, transcriptional activation by an ABF1 binding site upstream activation sequence is completely defective in an abf1(Ts) strain at the semipermissive temperature. These phenotypes provide evidence that the same protein, ABF1, functions in the initiation of DNA replication and transcriptional activation.


Item Type:Article
Additional Information:Copyright © 1992 by the American Society for Microbiology. Received 22 August 1991;accepted 22 November 1991. We thank Martin Budd for helpful discussions on generating temperature-sensitive mutants. We are grateful to Shlomo Eisenberg, Mitchell Smith, Scott Moye-Rowley, Carl Parker, Ambrose Jong, Martin Budd, and Karen Sitney for providing plasmids and strains used in this study. We also thank Shelley Diamond and Pat Koen for expert assistance in the flow cytometric analysis and Karen Oegema for assistance in cloning the 3' ABFI deletions. This work was supported by USPHS grant GM25508, a Procter and Gamble postdoctoral fellowship to P.R.R., and NIH training grant 5-T32-GM07616 (S.E.).
Subject Keywords:UPSTREAM ACTIVATING SEQUENCES; RECOGNIZE SPECIFIC SEQUENCES; YEAST PROTEIN; POLYMERASE-I; S-CEREVISIAE; SILENCER; GENE; PURIFICATION; ELEMENTS; MUTATIONS
Record Number:CaltechAUTHORS:RHOmcb92
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:RHOmcb92
Alternative URL:http://mcb.asm.org/cgi/content/abstract/12/3/1064
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:6011
Collection:CaltechAUTHORS
Deposited By: Lindsay Cleary
Deposited On:14 Nov 2006
Last Modified:26 Dec 2012 09:17

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