The Saccharomyces cerevisiae SPT8 Gene Encodes a Very Acidic Protein That Is Functionally Related to SPT3 and TATA-Binding Protein

INVESTIGATIONS

The Saccharomyces cerevisiae SPT8 Gene Encodes a Very Acidic Protein That Is Functionally Related to SPT3 and TATA-Binding Protein

D. M. Eisenmann, C. Chapon, S. M. Roberts, C. Dollard and F. Winston
Present address: Department of Developmental Biology, Stanford University School of Medicine, Stanford, California 94305.

Mutations in the Saccharomyces cerevisiae SPT8 gene were previously isolated as suppressors of retrotransposon insertion mutations in the 5' regions of the HIS4 and LYS2 genes. Mutations in SPT8 confer phenotypes similar to those caused by particular mutations in SPT15, which encodes the TATA-binding protein (TBP). These phenotypes are also similar to those caused by mutations in the SPT3 gene, which encodes a protein that directly interacts with TBP. We have now cloned and sequenced the SPT8 gene and have shown that it encodes a predicted protein of 602 amino acids with an extremely acidic amino terminus. In addition, the predicted SPT8 amino acid sequence contains one copy of a sequence motif found in multiple copies in a number of other eukaryotic proteins, including the {beta} subunit of heterotrimeric G proteins. To investigate further the relationship between SPT8, SPT3 and TBP, we have analyzed the effect of an spt8 null mutation in combination with different spt3 and spt15 mutations. This genetic analysis has shown that an spt8 deletion mutation is suppressed by particular spt3 alleles. Taken together with previous results, these data suggest that the SPT8 protein is required, directly or indirectly, for TBP function at particular promoters and that the role of SPT8 may be to promote a functional interaction between SPT3 and TBP.

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				J. A. Martens, P.-Y. J. Wu, and F. Winston Regulation of an intergenic transcript controls adjacent gene transcription in Saccharomyces cerevisiae Genes & Dev., November 15, 2005; 19(22): 2695 - 2704. [Abstract] [Full Text] [PDF]

				S. Jacobson and L. Pillus Molecular Requirements for Gene Expression Mediated by Targeted Histone Acetyltransferases Mol. Cell. Biol., July 1, 2004; 24(13): 6029 - 6039. [Abstract] [Full Text] [PDF]

				L. Warfield, J. A. Ranish, and S. Hahn Positive and negative functions of the SAGA complex mediated through interaction of Spt8 with TBP and the N-terminal domain of TFIIA Genes & Dev., May 1, 2004; 18(9): 1022 - 1034. [Abstract] [Full Text] [PDF]

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				D. E. Sterner and S. L. Berger Acetylation of Histones and Transcription-Related Factors Microbiol. Mol. Biol. Rev., June 1, 2000; 64(2): 435 - 459. [Abstract] [Full Text] [PDF]

				R. Belotserkovskaya, D. E. Sterner, M. Deng, M. H. Sayre, P. M. Lieberman, and S. L. Berger Inhibition of TATA-Binding Protein Function by SAGA Subunits Spt3 and Spt8 at Gcn4-Activated Promoters Mol. Cell. Biol., January 15, 2000; 20(2): 634 - 647. [Abstract] [Full Text]

				A. M. Dudley, C. Rougeulle, and F. Winston The Spt components of SAGA facilitate TBP binding to a promoter at a post-activator-binding step in vivo Genes & Dev., November 15, 1999; 13(22): 2940 - 2945. [Abstract] [Full Text]

				D. Donze, C. R. Adams, J. Rine, and R. T. Kamakaka The boundaries of the silenced HMR domain in Saccharomyces cerevisiae Genes & Dev., March 15, 1999; 13(6): 698 - 708. [Abstract] [Full Text]

				D. E. Sterner, P. A. Grant, S. M. Roberts, L. J. Duggan, R. Belotserkovskaya, L. A. Pacella, F. Winston, J. L. Workman, and S. L. Berger Functional Organization of the Yeast SAGA Complex: Distinct Components Involved in Structural Integrity, Nucleosome Acetylation, and TATA-Binding Protein Interaction Mol. Cell. Biol., January 1, 1999; 19(1): 86 - 98. [Abstract] [Full Text] [PDF]

				T. I. Lee and R. A. Young Regulation of gene expression by TBP-associated proteins Genes & Dev., May 15, 1998; 12(10): 1398 - 1408. [Full Text]

				E. vom Baur, M. Harbers, S.-J. Um, A. Benecke, P. Chambon, and R. Losson The yeast Ada complex mediates the ligand-dependent activation function AF-2 of retinoid X and estrogen receptors Genes & Dev., May 1, 1998; 12(9): 1278 - 1289. [Abstract] [Full Text]

				J. M. Madison, A. M. Dudley, and F. Winston Identification and Analysis of Mot3, a Zinc Finger Protein That Binds to the Retrotransposon Ty Long Terminal Repeat (delta ) in Saccharomyces cerevisiae Mol. Cell. Biol., April 1, 1998; 18(4): 1879 - 1890. [Abstract] [Full Text]

				L. Wang, L. Liu, and S. L. Berger Critical residues for histone acetylation by Gcn5, functioning in Ada and SAGA complexes, are also required for transcriptional function in vivo Genes & Dev., March 1, 1998; 12(5): 640 - 653. [Abstract] [Full Text]

				P A Grant, L Duggan, J Cote, S M Roberts, J E Brownell, R Candau, R Ohba, T Owen-Hughes, C D Allis, F Winston, et al. Yeast Gcn5 functions in two multisubunit complexes to acetylate nucleosomal histones: characterization of an Ada complex and the SAGA (Spt/Ada) complex. Genes & Dev., July 1, 1997; 11(13): 1640 - 1650. [Abstract] [PDF]