Our long-range goal is to understand mechanisms of negative regulation of transcription of eukaryotic genes. Our experiments will be performed in yeast, drosophila and in mammalian cells. We will isolate and analyze new negative regulators that work """"""""at a distance""""""""; these negative regulators bear """"""""repressing regions"""""""" fused to a DNA-binding fragment. We will study the mechanism by which the yeast negative regulators GAL80 inhibits activation by the yeast transcriptional activator GAL4, and we will study the effects of galactose (an inducer) on GAL80 action. We will analyze the ability of the yeast protein MCM1 to interact with either of two partners - 1 or 2 - to produce in one case a repressing and in the other case an activating complex, and we will attempt to isolate mammalian homologues of yeast proteins required for repression by MCM1/2.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM022526-18
Application #
2173945
Study Section
Molecular Biology Study Section (MBY)
Project Start
1978-09-01
Project End
1996-11-30
Budget Start
1993-12-01
Budget End
1994-11-30
Support Year
18
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Harvard University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
071723621
City
Cambridge
State
MA
Country
United States
Zip Code
02138
Sigal, G B; Bamdad, C; Barberis, A et al. (1996) A self-assembled monolayer for the binding and study of histidine-tagged proteins by surface plasmon resonance. Anal Chem 68:490-7
Astromoff, A; Ptashne, M (1995) A variant of lambda repressor with an altered pattern of cooperative binding to DNA sites. Proc Natl Acad Sci U S A 92:8110-4
Ohashi, Y; Brickman, J M; Furman, E et al. (1994) Modulating the potency of an activator in a yeast in vitro transcription system. Mol Cell Biol 14:2731-9
Reece, R J; Rickles, R J; Ptashne, M (1993) Overproduction and single-step purification of GAL4 fusion proteins from Escherichia coli. Gene 126:105-7
Bushman, F D; Shang, C; Ptashne, M (1989) A single glutamic acid residue plays a key role in the transcriptional activation function of lambda repressor. Cell 58:1163-71
Bushman, F D; Ptashne, M (1988) Turning lambda Cro into a transcriptional activator. Cell 54:191-7
Irwin, N; Ptashne, M (1987) Mutants of the catabolite activator protein of Escherichia coli that are specifically deficient in the gene-activation function. Proc Natl Acad Sci U S A 84:8315-9
Ptashne, M (1986) Gene regulation by proteins acting nearby and at a distance. Nature 322:697-701
Bushman, F D; Ptashne, M (1986) Activation of transcription by the bacteriophage 434 repressor. Proc Natl Acad Sci U S A 83:9353-7
Hochschild, A; Douhan 3rd, J; Ptashne, M (1986) How lambda repressor and lambda Cro distinguish between OR1 and OR3. Cell 47:807-16

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