Genetic Evidence for Transcriptional Activation by the Yeast IME1 Gene Product

INVESTIGATIONS

Genetic Evidence for Transcriptional Activation by the Yeast IME1 Gene Product

H. E. Smith, S. E. Driscoll, RAL. Sia, H. E. Yuan and A. P. Mitchell
Institute of Cancer Research and Department of Microbiology, Columbia University, New York, New York 10032

IME1 is required in yeast for meiosis and for expression of IME2 and other early meiotic genes. IME1 is a 360-amino acid polypeptide with central and C-terminal tyrosine-rich regions. We report here that a fusion protein composed of the lexA DNA-binding domain and IME1 activates transcription in vivo of a reporter gene containing upstream lexA binding sites. Activation by the fusion protein shares several features with natural IME1 activity: both are dependent on the RIM11 gene product; both are impaired by the same ime1 missense mutations; both are restored by intragenic suppressors. The central tyrosine-rich region is sufficient to activate transcription when fused to lexA. Deletion of this putative activation domain results in a defective IME1 derivative. Function of the deletion derivative is restored by fusion to the acidic Herpesvirus VP16 activation domain. The C-terminal tyrosine-rich region is dispensable for transcriptional activation; rather it renders activation dependent upon starvation and RIM11. Immunofluorescence studies indicate that an IME1-lacZ fusion protein is concentrated in the nucleus. These observations are consistent with a model in which IME1 normally stimulates IME2 expression by providing a transcriptional activation domain at the IME2 5' regulatory region.

This article has been cited by other articles:

M. Ramirez and J. Ambrona
Construction of Sterile ime1{Delta}-Transgenic Saccharomyces cerevisiae Wine Yeasts Unable To Disseminate in Nature
Appl. Envir. Microbiol., April 1, 2008; 74(7): 2129 - 2134.
[Abstract] [Full Text] [PDF]

M. Arevalo-Rodriguez and J. Heitman
Cyclophilin A Is Localized to the Nucleus and Controls Meiosis in Saccharomyces cerevisiae
Eukaryot. Cell, January 1, 2005; 4(1): 17 - 29.
[Abstract] [Full Text] [PDF]

I. Rubin-Bejerano, S. Sagee, O. Friedman, L. Pnueli, and Y. Kassir
The In Vivo Activity of Ime1, the Key Transcriptional Activator of Meiosis-Specific Genes in Saccharomyces cerevisiae, Is Inhibited by the Cyclic AMP/Protein Kinase A Signal Pathway through the Glycogen Synthase Kinase 3-{beta} Homolog Rim11
Mol. Cell. Biol., August 15, 2004; 24(16): 6967 - 6979.
[Abstract] [Full Text] [PDF]

L. Pnueli, I. Edry, M. Cohen, and Y. Kassir
Glucose and Nitrogen Regulate the Switch from Histone Deacetylation to Acetylation for Expression of Early Meiosis-Specific Genes in Budding Yeast
Mol. Cell. Biol., June 15, 2004; 24(12): 5197 - 5208.
[Abstract] [Full Text] [PDF]

D. M. Clifford, S. M. Marinco, and G. S. Brush
The Meiosis-specific Protein Kinase Ime2 Directs Phosphorylation of Replication Protein A
J. Biol. Chem., February 13, 2004; 279(7): 6163 - 6170.
[Abstract] [Full Text] [PDF]

N. Colomina, Y. Liu, M. Aldea, and E. Gari
TOR Regulates the Subcellular Localization of Ime1, a Transcriptional Activator of Meiotic Development in Budding Yeast
Mol. Cell. Biol., October 15, 2003; 23(20): 7415 - 7424.
[Abstract] [Full Text] [PDF]

R. M. Williams, M. Primig, B. K. Washburn, E. A. Winzeler, M. Bellis, C. Sarrauste de Menthiere, R. W. Davis, and R. E. Esposito
The Ume6 regulon coordinates metabolic and meiotic gene expression in yeast
PNAS, October 15, 2002; 99(21): 13431 - 13436.
[Abstract] [Full Text] [PDF]

N. Guttmann-Raviv, S. Martin, and Y. Kassir
Ime2, a Meiosis-Specific Kinase in Yeast, Is Required for Destabilization of Its Transcriptional Activator, Ime1
Mol. Cell. Biol., April 1, 2002; 22(7): 2047 - 2056.
[Abstract] [Full Text] [PDF]

K. F. Cooper and R. Strich
Saccharomycescerevisiae C-Type Cyclin Ume3p/Srb11p Is Required for Efficient Induction and Execution of Meiotic Development
Eukaryot. Cell, February 1, 2002; 1(1): 66 - 74.
[Abstract] [Full Text] [PDF]

B. K. Washburn and R. E. Esposito
Identification of the Sin3-Binding Site in Ume6 Defines a Two-Step Process for Conversion of Ume6 from a Transcriptional Repressor to an Activator in Yeast
Mol. Cell. Biol., March 15, 2001; 21(6): 2057 - 2069.
[Abstract] [Full Text]

G. Shenhar and Y. Kassir
A Positive Regulator of Mitosis, Sok2, Functions as a Negative Regulator of Meiosis in Saccharomyces cerevisiae
Mol. Cell. Biol., March 1, 2001; 21(5): 1603 - 1612.
[Abstract] [Full Text]

Y. Xiao and A. P. Mitchell
Shared Roles of Yeast Glycogen Synthase Kinase 3 Family Members in Nitrogen-Responsive Phosphorylation of Meiotic Regulator Ume6p
Mol. Cell. Biol., August 1, 2000; 20(15): 5447 - 5453.
[Abstract] [Full Text]

D. Davis, R. B. Wilson, and A. P. Mitchell
RIM101-Dependent and -Independent Pathways Govern pH Responses in Candida albicans
Mol. Cell. Biol., February 1, 2000; 20(3): 971 - 978.
[Abstract] [Full Text]

K. Malathi, Y. Xiao, and A. P. Mitchell
Catalytic Roles of Yeast GSK3{beta}/Shaggy Homolog Rim11p in Meiotic Activation
Genetics, November 1, 1999; 153(3): 1145 - 1152.
[Abstract] [Full Text]

S. M. Honigberg and R. H. Lee
Snf1 Kinase Connects Nutritional Pathways Controlling Meiosis in Saccharomyces cerevisiae
Mol. Cell. Biol., August 1, 1998; 18(8): 4548 - 4555.
[Abstract] [Full Text]

S. Sagee, A. Sherman, G. Shenhar, K. Robzyk, N. Ben-Doy, G. Simchen, and Y. Kassir
Multiple and Distinct Activation and Repression Sequences Mediate the Regulated Transcription of IME1, a Transcriptional Activator of Meiosis-Specific Genes in Saccharomyces cerevisiae
Mol. Cell. Biol., April 1, 1998; 18(4): 1985 - 1995.
[Abstract] [Full Text]

H Friesen, R Lunz, S Doyle, and J Segall
Mutation of the SPS1-encoded protein kinase of Saccharomyces cerevisiae leads to defects in transcription and morphology during spore formation.
Genes & Dev., September 15, 1994; 8(18): 2162 - 2175.
[Abstract] [PDF]

R Strich, R T Surosky, C Steber, E Dubois, F Messenguy, and R E Esposito
UME6 is a key regulator of nitrogen repression and meiotic development.
Genes & Dev., April 1, 1994; 8(7): 796 - 810.
[Abstract] [PDF]

				M. Arevalo-Rodriguez and J. Heitman Cyclophilin A Is Localized to the Nucleus and Controls Meiosis in Saccharomyces cerevisiae Eukaryot. Cell, January 1, 2005; 4(1): 17 - 29. [Abstract] [Full Text] [PDF]

				I. Rubin-Bejerano, S. Sagee, O. Friedman, L. Pnueli, and Y. Kassir The In Vivo Activity of Ime1, the Key Transcriptional Activator of Meiosis-Specific Genes in Saccharomyces cerevisiae, Is Inhibited by the Cyclic AMP/Protein Kinase A Signal Pathway through the Glycogen Synthase Kinase 3-{beta} Homolog Rim11 Mol. Cell. Biol., August 15, 2004; 24(16): 6967 - 6979. [Abstract] [Full Text] [PDF]

				L. Pnueli, I. Edry, M. Cohen, and Y. Kassir Glucose and Nitrogen Regulate the Switch from Histone Deacetylation to Acetylation for Expression of Early Meiosis-Specific Genes in Budding Yeast Mol. Cell. Biol., June 15, 2004; 24(12): 5197 - 5208. [Abstract] [Full Text] [PDF]

				D. M. Clifford, S. M. Marinco, and G. S. Brush The Meiosis-specific Protein Kinase Ime2 Directs Phosphorylation of Replication Protein A J. Biol. Chem., February 13, 2004; 279(7): 6163 - 6170. [Abstract] [Full Text] [PDF]

				N. Colomina, Y. Liu, M. Aldea, and E. Gari TOR Regulates the Subcellular Localization of Ime1, a Transcriptional Activator of Meiotic Development in Budding Yeast Mol. Cell. Biol., October 15, 2003; 23(20): 7415 - 7424. [Abstract] [Full Text] [PDF]

				R. M. Williams, M. Primig, B. K. Washburn, E. A. Winzeler, M. Bellis, C. Sarrauste de Menthiere, R. W. Davis, and R. E. Esposito The Ume6 regulon coordinates metabolic and meiotic gene expression in yeast PNAS, October 15, 2002; 99(21): 13431 - 13436. [Abstract] [Full Text] [PDF]

				N. Guttmann-Raviv, S. Martin, and Y. Kassir Ime2, a Meiosis-Specific Kinase in Yeast, Is Required for Destabilization of Its Transcriptional Activator, Ime1 Mol. Cell. Biol., April 1, 2002; 22(7): 2047 - 2056. [Abstract] [Full Text] [PDF]

				K. F. Cooper and R. Strich Saccharomycescerevisiae C-Type Cyclin Ume3p/Srb11p Is Required for Efficient Induction and Execution of Meiotic Development Eukaryot. Cell, February 1, 2002; 1(1): 66 - 74. [Abstract] [Full Text] [PDF]

				B. K. Washburn and R. E. Esposito Identification of the Sin3-Binding Site in Ume6 Defines a Two-Step Process for Conversion of Ume6 from a Transcriptional Repressor to an Activator in Yeast Mol. Cell. Biol., March 15, 2001; 21(6): 2057 - 2069. [Abstract] [Full Text]

				G. Shenhar and Y. Kassir A Positive Regulator of Mitosis, Sok2, Functions as a Negative Regulator of Meiosis in Saccharomyces cerevisiae Mol. Cell. Biol., March 1, 2001; 21(5): 1603 - 1612. [Abstract] [Full Text]

				Y. Xiao and A. P. Mitchell Shared Roles of Yeast Glycogen Synthase Kinase 3 Family Members in Nitrogen-Responsive Phosphorylation of Meiotic Regulator Ume6p Mol. Cell. Biol., August 1, 2000; 20(15): 5447 - 5453. [Abstract] [Full Text]

				D. Davis, R. B. Wilson, and A. P. Mitchell RIM101-Dependent and -Independent Pathways Govern pH Responses in Candida albicans Mol. Cell. Biol., February 1, 2000; 20(3): 971 - 978. [Abstract] [Full Text]

				K. Malathi, Y. Xiao, and A. P. Mitchell Catalytic Roles of Yeast GSK3{beta}/Shaggy Homolog Rim11p in Meiotic Activation Genetics, November 1, 1999; 153(3): 1145 - 1152. [Abstract] [Full Text]

				S. M. Honigberg and R. H. Lee Snf1 Kinase Connects Nutritional Pathways Controlling Meiosis in Saccharomyces cerevisiae Mol. Cell. Biol., August 1, 1998; 18(8): 4548 - 4555. [Abstract] [Full Text]

				S. Sagee, A. Sherman, G. Shenhar, K. Robzyk, N. Ben-Doy, G. Simchen, and Y. Kassir Multiple and Distinct Activation and Repression Sequences Mediate the Regulated Transcription of IME1, a Transcriptional Activator of Meiosis-Specific Genes in Saccharomyces cerevisiae Mol. Cell. Biol., April 1, 1998; 18(4): 1985 - 1995. [Abstract] [Full Text]

				H Friesen, R Lunz, S Doyle, and J Segall Mutation of the SPS1-encoded protein kinase of Saccharomyces cerevisiae leads to defects in transcription and morphology during spore formation. Genes & Dev., September 15, 1994; 8(18): 2162 - 2175. [Abstract] [PDF]

				R Strich, R T Surosky, C Steber, E Dubois, F Messenguy, and R E Esposito UME6 is a key regulator of nitrogen repression and meiotic development. Genes & Dev., April 1, 1994; 8(7): 796 - 810. [Abstract] [PDF]