This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hagen, D C Articles by Sprague, G F Search for Related Content PubMed PubMed Citation Articles by Hagen, D C Articles by Sprague, G F, Jr

 Previous Article  |  Next Article 

Mol Cell Biol. 1993 November; 13(11): 6866-6875

Transcription of alpha-specific genes in Saccharomyces cerevisiae: DNA sequence requirements for activity of the coregulator alpha 1.

Institute of Molecular Biology, University of Oregon, Eugene 97403.

ABSTRACT

Transcription activation of alpha-specific genes in Saccharomyces cerevisiae is regulated by two proteins, MCM1 and alpha 1, which bind to DNA sequences, called P'Q elements, found upstream of alpha-specific genes. Neither MCM1 nor alpha 1 alone binds efficiently to P'Q elements. Together, however, they bind cooperatively in a manner that requires both the P' sequence, which is a weak binding site for MCM1, and the Q sequence, which has been postulated to be the binding site for alpha 1. We analyzed a collection of point mutations in the P'Q element of the STE3 gene to determine the importance of individual base pairs for alpha-specific gene transcription. Within the 10-bp conserved Q sequence, mutations at only three positions strongly affected transcription activation in vivo. These same mutations did not affect the weak binding to P'Q displayed by MCM1 alone. In vitro DNA binding assays showed a direct correlation between the ability of the mutant sequences to form ternary P'Q-MCM1-alpha 1 complexes and the degree to which transcription was activated in vivo. Thus, the ability of alpha 1 and MCM1 to bind cooperatively to P'Q elements is critical for activation of alpha-specific genes. In all natural alpha-specific genes the Q sequence is adjacent to the degenerate side of P'. To test the significance of this geometry, we created several novel juxtapositions of P, P', and Q sequences. When the Q sequence was opposite the degenerate side, the composite QP' element was inactive as a promoter element in vivo and unable to form stable ternary QP'-MCM1-alpha 1 complexes in vitro. We also found that addition of a Q sequence to a strong MCM1 binding site allows the addition of alpha 1 to the complex. This finding, together with the observation that Q-element point mutations affected ternary complex formation but not the weak binding of MCM1 alone, supports the idea that the Q sequence serves as a binding site for alpha 1.

This article has been cited by other articles:

• Zill, O. A., Rine, J. (2008). Interspecies variation reveals a conserved repressor of {alpha}-specific genes in Saccharomyces yeasts. Genes Dev. 22: 1704-1716 [Abstract] [Full Text]  
• Dalgaard, J. Z., Vengrova, S. (2004). Selective Gene Expression in Multigene Families from Yeast to Mammals. Sci Signal 2004: re17-re17 [Abstract] [Full Text]  
• Carr, E. A., Mead, J., Vershon, A. K. (2004). {alpha}1-induced DNA bending is required for transcriptional activation by the Mcm1-{alpha}1 complex. Nucleic Acids Res 32: 2298-2305 [Abstract] [Full Text]  
• Mead, J., Bruning, A. R., Gill, M. K., Steiner, A. M., Acton, T. B., Vershon, A. K. (2002). Interactions of the Mcm1 MADS Box Protein with Cofactors That Regulate Mating in Yeast. Mol. Cell. Biol. 22: 4607-4621 [Abstract] [Full Text]  
• Zhong, H., McCord, R., Vershon, A. K. (1999). Identification of Target Sites of the alpha 2-Mcm1 Repressor Complex in the Yeast Genome. Genome Res 9: 1040-1047 [Abstract] [Full Text]  
• Partridge, J. F., Mikesell, G. E., Breeden, L. L. (1997). Cell Cycle-dependent Transcription of CLN1 Involves Swi4 Binding to MCB-like Elements. J. Biol. Chem. 272: 9071-9077 [Abstract] [Full Text]