Isolation and Characterization of Mutants Constitutive for Expression of the fbp1 Gene of Schizosaccharomyces pombe

INVESTIGATIONS

Isolation and Characterization of Mutants Constitutive for Expression of the fbp1 Gene of Schizosaccharomyces pombe

C. S. Hoffman and F. Winston
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115

Transcription of the fbp1 gene of Schizosaccharomyces pombe, encoding fructose-1,6-bisphosphatase, is glucose repressible. We have constructed two hybrid genes, containing the fbp1 promoter, that allow selection for mutations that alter transcriptional regulation of fbp1. Strains carrying fbp1-ura4 and fbp1-lacZ fusions are phenotypically Ura(-), resistant to 5-fluoro-orotic acid, and express a low level of {beta}-galactosidase activity when grown under repressing conditions (8% glucose). By selecting for Ura(+) strains grown under repressing conditions, we have isolated 187 independent mutants that constitutively express the fbp1-ura4 fusion. These mutants identify ten complementation groups that represent ten unlinked git (glucose insensitive transcription) genes. The git gene products are required in trans for glucose repression of expression from the fbp1 promoter since these mutations also alter expression of the fbp1-lacZ fusion. We have shown that transcription of the wild type fbp1 gene in most git mutants is elevated to a level consistent with the increased expression of the fbp1-lacZ hybrid gene. Mutations in some git genes confer additional phenotypes such as slow growth, temperature-sensitive lethality and reduced spore viability. Therefore, some of these genes are likely to encode factors that are of general importance for S. pombe transcription.

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				R. S. Kao, E. Morreale, L. Wang, F. D. Ivey, and C. S. Hoffman Schizosaccharomyces pombe Git1 Is a C2-Domain Protein Required for Glucose Activation of Adenylate Cyclase Genetics, May 1, 2006; 173(1): 49 - 61. [Abstract] [Full Text] [PDF]

				L. Wang, K. Griffiths Jr., Y. H. Zhang, F. D. Ivey, and C. S. Hoffman Schizosaccharomyces pombe Adenylate Cyclase Suppressor Mutations Suggest a Role for cAMP Phosphodiesterase Regulation in Feedback Control of Glucose/cAMP Signaling Genetics, December 1, 2005; 171(4): 1523 - 1533. [Abstract] [Full Text] [PDF]

				F. D. Ivey and C. S. Hoffman Direct activation of fission yeast adenylate cyclase by the Gpa2 G{alpha} of the glucose signaling pathway PNAS, April 26, 2005; 102(17): 6108 - 6113. [Abstract] [Full Text] [PDF]

				C. S. Hoffman Except in Every Detail: Comparing and Contrasting G-Protein Signaling in Saccharomyces cerevisiae and Schizosaccharomyces pombe Eukaryot. Cell, March 1, 2005; 4(3): 495 - 503. [Full Text] [PDF]

				M. Madrid, T. Soto, A. Franco, V. Paredes, J. Vicente, E. Hidalgo, M. Gacto, and J. Cansado A Cooperative Role for Atf1 and Pap1 in the Detoxification of the Oxidative Stress Induced by Glucose Deprivation in Schizosaccharomyces pombe J. Biol. Chem., October 1, 2004; 279(40): 41594 - 41602. [Abstract] [Full Text] [PDF]

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				K. Hirota, C. S. Hoffman, T. Shibata, and K. Ohta Fission Yeast Tup1-Like Repressors Repress Chromatin Remodeling at the fbp1+ Promoter and the ade6-M26 Recombination Hotspot Genetics, October 1, 2003; 165(2): 505 - 515. [Abstract] [Full Text] [PDF]

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				L. Taricani, M. L. Tejada, and P. G. Young The Fission Yeast ES2 Homologue, Bis1, Interacts with the Ish1 Stress-responsive Nuclear Envelope Protein J. Biol. Chem., March 15, 2002; 277(12): 10562 - 10572. [Abstract] [Full Text] [PDF]

				T. Higuchi, Y. Watanabe, and M. Yamamoto Protein Kinase A Regulates Sexual Development and Gluconeogenesis through Phosphorylation of the Zn Finger Transcriptional Activator Rst2p in Fission Yeast Mol. Cell. Biol., January 1, 2002; 22(1): 1 - 11. [Abstract] [Full Text] [PDF]

				S. Landry and C. S. Hoffman The git5 G{beta} and git11 G{{gamma}} Form an Atypical G{beta}{{gamma}} Dimer Acting in the Fission Yeast Glucose/cAMP Pathway Genetics, March 1, 2001; 157(3): 1159 - 1168. [Abstract] [Full Text]

				R. T. K. Janoo, L. A. Neely, B. R. Braun, S. K. Whitehall, and C. S. Hoffman Transcriptional Regulators of the Schizosaccharomyces pombe fbp1 Gene Include Two Redundant Tup1p-like Corepressors and the CCAAT Binding Factor Activation Complex Genetics, March 1, 2001; 157(3): 1205 - 1215. [Abstract] [Full Text]

				K. B. Lengeler, R. C. Davidson, C. D'souza, T. Harashima, W.-C. Shen, P. Wang, X. Pan, M. Waugh, and J. Heitman Signal Transduction Cascades Regulating Fungal Development and Virulence Microbiol. Mol. Biol. Rev., December 1, 2000; 64(4): 746 - 785. [Abstract] [Full Text] [PDF]

				C.-R. Chen, J. Chen, and E. C. Chang A Conserved Interaction between Moe1 and Mal3 Is Important for Proper Spindle Formation in Schizosaccharomyces pombe Mol. Biol. Cell, December 1, 2000; 11(12): 4067 - 4077. [Abstract] [Full Text]

				R. M. Welton and C. S. Hoffman Glucose Monitoring in Fission Yeast via the gpa2 G{alpha}, the git5 G{beta} and the git3 Putative Glucose Receptor Genetics, October 1, 2000; 156(2): 513 - 521. [Abstract] [Full Text]

				L. A. Neely and C. S. Hoffman Protein Kinase A and Mitogen-Activated Protein Kinase Pathways Antagonistically Regulate Fission Yeast fbp1 Transcription by Employing Different Modes of Action at Two Upstream Activation Sites Mol. Cell. Biol., September 1, 2000; 20(17): 6426 - 6434. [Abstract] [Full Text]

				S. Landry, M. T. Pettit, E. Apolinario, and C. S. Hoffman The Fission Yeast git5 Gene Encodes a G{beta} Subunit Required for Glucose-Triggered Adenylate Cyclase Activation Genetics, April 1, 2000; 154(4): 1463 - 1471. [Abstract] [Full Text]

				M. C. Lorenz, X. Pan, T. Harashima, M. E. Cardenas, Y. Xue, J. P. Hirsch, and J. Heitman The G Protein-Coupled Receptor Gpr1 Is a Nutrient Sensor That Regulates Pseudohyphal Differentiation in Saccharomyces cerevisiae Genetics, February 1, 2000; 154(2): 609 - 622. [Abstract] [Full Text]

				J. M. Gancedo Yeast Carbon Catabolite Repression Microbiol. Mol. Biol. Rev., June 1, 1998; 62(2): 334 - 361. [Abstract] [Full Text] [PDF]

				M.-Y. Chen, Y. Long, and P. N. Devreotes A novel cytosolic regulator, Pianissimo, is required for chemoattractant receptor and G protein-mediated activation of the 12 transmembrane domain adenylyl cyclase in Dictyostelium Genes & Dev., December 1, 1997; 11(23): 3218 - 3231. [Abstract] [Full Text] [PDF]

				Z Hao, A Furunobu, A Nagata, and H Okayama A zinc finger protein required for stationary phase viability in fission yeast J. Cell Sci., January 10, 1997; 110(20): 2557 - 2566. [Abstract] [PDF]

				T Caspari Onset of gluconate-H+ symport in Schizosaccharomyces pombe is regulated by the kinases Wis1 and Pka1, and requires the gti1+ gene product J. Cell Sci., January 10, 1997; 110(20): 2599 - 2608. [Abstract] [PDF]

				P Dal Santo, B Blanchard, and C. Hoffman The Schizosaccharomyces pombe pyp1 protein tyrosine phosphatase negatively regulates nutrient monitoring pathways J. Cell Sci., January 7, 1996; 109(7): 1919 - 1925. [Abstract] [PDF]

				S Stettler, E Warbrick, S Prochnik, S Mackie, and P Fantes The wis1 signal transduction pathway is required for expression of cAMP-repressed genes in fission yeast J. Cell Sci., January 7, 1996; 109(7): 1927 - 1935. [Abstract] [PDF]

				S. Byrne and C. Hoffman Six git genes encode a glucose-induced adenylate cyclase activation pathway in the fission yeast Schizosaccharomyces pombe J. Cell Sci., January 8, 1993; 105(4): 1095 - 1100. [Abstract] [PDF]

				C S Hoffman and F Winston Glucose repression of transcription of the Schizosaccharomyces pombe fbp1 gene occurs by a cAMP signaling pathway. Genes & Dev., April 1, 1991; 5(4): 561 - 571. [Abstract] [PDF]