Deletion analysis of GAL4 defines two transcriptional activating segments

We describe the activities of a wide array of deletion mutants of GAL4, a yeast transcriptional activator. We identify two short regions of GAL4, each of which activates transcription when fused to the DNA-binding region of the molecule. Very large portions of GAL4 are not required for gene activation.

[1]  L. Guarente,et al.  A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[2]  L. Guarente,et al.  Fusion of Escherichia coli lacZ to the cytochrome c gene of Saccharomyces cerevisiae. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[3]  S. Johnston,et al.  Analysis of the Kluyveromyces lactis positive regulatory gene LAC9 reveals functional homology to, but sequence divergence from, the Saccharomyces cerevisiae GAL4 gene. , 1986, Nucleic acids research.

[4]  A. Laughon,et al.  Isolation and preliminary characterization of the GAL4 gene, a positive regulator of transcription in yeast. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[5]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[6]  K. Murata,et al.  Transformation of intact yeast cells treated with alkali cations , 1983 .

[7]  R. W. Davis,et al.  Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae , 1984, Molecular and cellular biology.

[8]  M. Ptashne,et al.  Saccharomyces cerevisiae GAL1-GAL10 divergent promoter region: location and function of the upstream activating sequence UASG , 1984, Molecular and cellular biology.

[9]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[10]  Mark Ptashne,et al.  Gene regulation by proteins acting nearby and at a distance , 1986, Nature.

[11]  S. Johnston,et al.  Isolation of the yeast regulatory gene GAL4 and analysis of its dosage effects on the galactose/melibiose regulon. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[12]  W. Gilbert,et al.  Sequencing end-labeled DNA with base-specific chemical cleavages. , 1980, Methods in enzymology.

[13]  A. Laughon,et al.  Primary structure of the Saccharomyces cerevisiae GAL4 gene , 1984, Molecular and cellular biology.

[14]  K. Matsumoto,et al.  Isolation and characterization of dominant mutations resistant to carbon catabolite repression of galactokinase synthesis in Saccharomyces cerevisiae , 1981, Molecular and cellular biology.

[15]  K. Struhl,et al.  Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of Yeast , 1986, Cell.

[16]  David Botstein,et al.  Two differentially regulated mRNAs with different 5′ ends encode secreted and intracellular forms of yeast invertase , 1982, Cell.

[17]  M. Ptashne,et al.  Use of lacZ fusions to delimit regulatory elements of the inducible divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae , 1984, Molecular and cellular biology.

[18]  Michael Shales,et al.  Extensive homology among the largest subunits of eukaryotic and prokaryotic RNA polymerases , 1985, Cell.

[19]  J. Biggs,et al.  Structure of the eukaryotic transcription apparatus: Features of the gene for the largest subunit of Drosophila RNA polymerase II , 1985, Cell.

[20]  Michael R. Green,et al.  Normal and mutant human β-globin pre-mRNAs are faithfully and efficiently spliced in vitro , 1984, Cell.

[21]  R. Kornberg,et al.  A GAL family of upstream activating sequences in yeast: roles in both induction and repression of transcription. , 1986, The EMBO journal.

[22]  K. Yamamoto,et al.  Glucocorticoid receptor mutants that are constitutive activators of transcriptional enhancement , 1987, Nature.

[23]  R. Brent,et al.  A eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor , 1985, Cell.

[24]  R. Kornberg,et al.  Specific protein binding to far upstream activating sequences in polymerase II promoters. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[25]  R. Kornberg,et al.  A region flanking the GAL7 gene and a binding site for GAL4 protein as upstream activating sequences in yeast. , 1985, Journal of molecular biology.

[26]  B. G. Adams.  Induction of Galactokinase in Saccharomyces cerevisiae: Kinetics of Induction and Glucose Effects , 1972, Journal of bacteriology.

[27]  H. Blumberg,et al.  Sequence homology of the yeast regulatory protein ADR1 with Xenopus transcription factor TFIIIA , 1986, Nature.

[28]  M. Ptashne,et al.  Specific DNA binding of GAL4, a positive regulatory protein of yeast , 1985, Cell.

[29]  K. Murata,et al.  Transformation of intact yeast cells treated with alkali cations. , 1984, Journal of bacteriology.

[30]  P. Farabaugh.  Sequence of the lacI gene , 1978, Nature.

[31]  M. Ptashne.  A Genetic Switch , 1986 .

[32]  J. Bennetzen,et al.  The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase. , 1982, The Journal of biological chemistry.

[33]  M. Ptashne,et al.  Repressor structure and the mechanism of positive control , 1983, Cell.

[34]  P. Silver,et al.  Amino terminus of the yeast GAL4 gene product is sufficient for nuclear localization. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[35]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[36]  M. Ptashne,et al.  Separation of DNA binding from the transcription-activating function of a eukaryotic regulatory protein. , 1986, Science.

[37]  K. Matsumoto,et al.  [Regulatory circuits for gene expression: the metabolism of galactose and phosphate in Saccharomyces cerevisiae]. , 1984, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.