Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Reexamination Certificate
2002-10-09
2004-10-05
Saidha, Tekchand (Department: 1652)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C435S183000, C435S232000, C435S252300, C435S320100, C536S023200
Reexamination Certificate
active
06800459
ABSTRACT:
FIELD OF THE INVENTION
The invention relates generally to plant molecular biology. In particular, the invention relates to compositions and methods for the regulation of plant growth and development through the modulation of either chorismate synthase or chorismate mutase gene expression or activity.
BACKGROUND OF THE INVENTION
Chorismate is an essential substrate for the synthesis of p-aminobenzoate, folate, ubiquinone and the aromatic amino acids tryptophan, phenylalanine and tyrosine. Chorismate is produced in the seventh step of the shikimate pathway. This pathway has been described in numerous publications. See, for example,
Shikimic Acid: Metabolism and Metabolites
, John Wiley & Sons, Winchester, UK, 1993. The first four steps of the shikimate biosynthetic pathway lead to the production of shikimate. Shikimate is then converted to chorismate in next three steps of the pathway. First, shikimate is converted to shikimate-5-phosphate by shikimate kinase. Next, 3-enolpyruvateshikimate 5-phosphate synthase converts shikimate-5-phosphate to 5-enolpyruvylshikimate 3-phosphate, which is then converted to chorismate by the enzyme chrosimate synthase.
The pathway leading to aromatic amino acid synthesis branches at chorismate. One branch leads to the synthesis of tryptophan. The other branch leads to the synthesis of phenylalanine and tyrosine. Thus, chorismate is the last common intermediate in the synthesis of tryptophan, phenylalanine and tyrosine.
In the branch leading to phenylalanine and tyrosine synthesis, chorismate is converted to prephenate by the enzyme chorismate mutase. Prephenate is the last common intermediate for biosyntheses of phenylalanine and tyrosine by two independent pathways that are present in both eukaryotes and prokaryotes.
Three isozymes of chorismate mutase, CM-1, CM-2 and CM-3 have been found in plants. Mobley et al. (1999)
Gene
240:115-123. CM-1 and CM-3 are plastidic, while CM-2 is cytosolic. In
Arabidopsis thaliana
, CM-1 has 53% amino acid similarity with CM-2 and 68% amino acid similarity with CM-3.
The conversion of shikimate-5-phosphate to 5-enolpyruvylshikimate 3-phosphate is blocked by the commercially successful herbicide Roundup™ (glyphosate). Accordingly, the shikimate pathway has been considered an attractive target for herbicides (PCT publication WO 00/05353, the contents of which are incorporated by reference, and Roberts et al. (1998)
Nature
393:801-805). However, while it has been suggested that that chorismate synthase could be a candidate for a herbicide target (Bomemann et al. (1985)
J Biol Chem
270:228111-22815), previous studies have not been able to ascertain whether chorismate synthase or chorismate mutase are essential for plant growth, which is a key parameter for determining potential herbicide targets. Nor are there any herbicides that are known to act by modifying the activity of either of these enzymes. Thus, it is necessary to determine whether such enzymes are critical to plant growth, before they can be considered useful targets in assays for the identification of herbicides and herbicide candidates.
SUMMARY OF THE INVENTION
The present inventors have discovered that chorismate mutase and chorismate synthase are essential for plant growth. Specifically, the inhibition of chorismate mutase or chorismate synthase gene expression in plant seedlings results in severe chlorosis, reduced growth and developmental abnormalities. Thus, in one aspect, the present invention provides compositions for the modulation of plant growth or development comprising chorismate synthase and chorismate mutase antisense and sense polynucleotides, dsRNA and ribozymes, and related expression cassettes and vectors. The compositions of the invention are particularly useful for the modulation and inhibition of plant growth. The invention further provides plants, plant cells, and seeds containing the polynucleotides of the invention.
The inventors have proven that chorismate synthase and chorismate mutase can be used as targets for the identification of herbicides. Thus, the present invention also provides methods for the identification of chemicals that modulate chorismate synthase and chorismate mutase biochemical reactions. The methods of the invention are useful for the identification of herbicides and for the inhibition of plant growth and development. In addition, the methods of the invention are useful for the identification of compounds that stimulate the expression or function of chorismate synthase or chorismate mutase expression or function. Such compounds can be used to promote or manipulate plant growth and development.
REFERENCES:
patent: 4753883 (1988-06-01), Backman et al.
patent: 5120837 (1992-06-01), Fotheringham et al.
patent: 5187071 (1993-02-01), Fischer et al.
patent: 5776736 (1998-07-01), Frost et al.
patent: 5795715 (1998-08-01), Livache et al.
patent: 5948612 (1999-09-01), Bascomb et al.
patent: 96/19103 (1996-06-01), None
patent: 98/59061 (1998-12-01), None
patent: 00/05353 (2000-02-01), None
patent: 00/16609 (2000-03-01), None
patent: 00/31281 (2000-06-01), None
Bornemann, Stephen et al., “Escherichia colichorismate synthase: a deuterium kinetic-isotope effect under a single-turnover and steady-state conditions shows that a flavin intermediate forms before the C-(6proR)-H bond is cleaved”, Biochem J., vol. 305, pp. 707-710 (1995).
Braun, Martin et al., Enzymatic properties of chorismate synthase isozymes of tomato (Lycopersicon esculentumMill.). Planta, vol. 200 pp. 64-70 (1996).
Cotton, R.G.H. et al., “The biosynthesis of phenylalanine and tyrosine in the pea (Pisum sativam): Chorismate mutase”. Biochimica et Biophysica Acta, vol. 156, pp. 187-189, (1968).
Eberhard, Jenny et al., “Isolation of a cDNA from tomato coding for an unregulated, cytosolic chorismate mutase”, Plant Molecular Biology, vol. 31, pp. 917-922, (1996).
Gilchrist, D.G. et al., “Chorismate Mutase from Mung Bean and Sorghum”, Methods in Enzymology, vol. 142, [54] pp. 450-463, 1987).
Gilchrist, D.G. et al., “Regulation of Aromatic Amino Acid Biosynthesis in Higher Plants, Properties of an Aromatic Amino Acid-Sensitive Chorismate Mutase (CM-1) from Mung Bean”, Archives of Biochemistry and Biophysics, vol. 164, pp. 95-105, (1974).
Gilchrist, D. G., “Regulation of Aromatic Amino Acid Biosynthesis in Higher Plants, Properties of an Aromatics Amino Acid Insensitive Chorismate Mutase (CM-2) from Mung Bean”, Archives of Biochemistry of Biophysics, vol. 171, pp. 36-42, (1975).
Gorisch, Helmut, “Chorismate Mutase fromStreptomyces aureofaciens”, Methods in Enzymology, vol. 142, [55] pp. 463-472, (1987).
Gorisch, Helmut, “A New Test for Chorismate Mutase Acitivity”, Analytical Biochemistry, vol. 86, pp. 764-768, (1978).
Henstrand, John M. et al., “Cloning and Characterization of a Heterologously Expressed Bifunctional Chorismate Synthase/Flavin Reductase fromNeurospora crassa”, The Journal of Biological Chemistry, vol. 270, No. 35, pp. 20447-20452, (1995).
Krappmann, Sven et al., The aroC Gene ofAspergillus nidulansCodes for a Monofunctional, Allosterically Regulated Chorismate Mutase, The Journal of Biological Chemistry, vol. 274, No. 32, pp. 22275-22282, (1999).
Kuroki, Gary W. et al., Purification and Characterization of an Inducible Aromatic Amino Acid-Sensitive Form of Chorismate Mutase fromSolanum tuberosumL. Tubers, vol. 260, No. 2, pp. 616-621, (1988).
Macheroux, Peter et al., Studies with Flavin Analogs Provide Evidence That a Protonated Reduced FMN Is the Substrate-induced Transient Intermediate in the Reaction ofEscherichia coliChorismate Synthase, The Journal of Biological Chemistry, vol. 271, No. 42, pp. 25850-25858, (1996).
Ramjee, Manoj K. et al., “A Continuous, Anaerobic Spectrophotometric Assay for Chorismate Synthase Activity That Utilizes Photoreduced Flavin Mononucelotide”, Analytical Biochemistry, vol. 220, pp. 137-141 (1994).
Schmidheini, Tobias, et al., “A Single Point Mutation Results in a Constitutively Activated And Feedback-Resistant Chorismate Mutase ofSaccharomyces cerevisiase”, Journal of Bacteriology, vol
Ascenzi Robert A.
Boyes Douglas C.
Davis Keith R.
Görlach Jörn
Hamilton Carol M.
Hofmeyer Timothy G.
Kiefer Laura L.
Paradigm Genetics, Inc.
Saidha Tekchand
Spencer Deborah H.
LandOfFree
Methods and compositions for the modulation of chorismate... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Methods and compositions for the modulation of chorismate..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Methods and compositions for the modulation of chorismate... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3266060