Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Transferase other than ribonuclease
Patent
1995-08-02
1999-06-15
Wax, Robert A.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Transferase other than ribonuclease
4353201, 435410, 536 232, 935 22, C07H 2104, C12N 1500, C12N 912, C12N 500
Patent
active
059121564
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a novel polypeptide having cold-stable pyruvate, orthophosphate dikinase (hereinafter also referred to as "PPDK") activity, a cloned DNA encoding the same and a recombinant vector containing the DNA, as means for giving cold-stability to plants. The present invention also relates to plants transformed with the DNA according to the present invention.
BACKGROUND ART
C.sub.4 plants have high abilities of photosynthesis under the conditions of strong light, high temperature or low CO.sub.2. However, their photosynthesis abilities are largely reduced under low temperature except for those which are adapted to low temperature conditions. Although PPDK (EC 2.7.9.1, that catalyzes the reaction in which AMP, phosphoenol pyruvate and pyrophosphate are produced from ATP, pyruvate and orthophosphate) is one of the important enzymes in C.sub.4 path, its activity is not sufficient with respect to the photosynthesis rate in leaf tissue, so that it is one of the enzymes which determine the rate of CO.sub.2 fixation in C.sub.4 photosynthesis. Further, it has been pointed out simultaneously with the discovery of this enzyme that this enzyme is cold-sensitive. In case of maize PPDK, the enzyme activity has a point of inflection at 11.7.degree. C. This temperature is coincident with the limit temperature of growth of maize. From these, it is thought that PPDK is one of the causes which reduce the photosynthesis rate of C.sub.4 plants at low temperature. Therefore, by improving the cold-sensitivity of PPDK, the limit temperature of growth of maize may be lowered. Flaveria brownii which is a plant belonging to the family Compositae is classified into C.sub.3 /C.sub.4 intermediate type, and it is known that its PPDK is not substantially inactivated by low temperature treatment at 0.degree. C. (Burnell JN: A comparative study of the cold-sensitivity of pyruvate, Pi dikinase in Flaveria species. Plant Cell Physiol. 31, 295-297 (1990)).
By cloning the gene encoding the cold-stable PPDK of Flaveria brownii, and by transforming a plant with the gene, it is expected that resistance to coldness can be given to the plant.
DISCLOSURE OF THE INVENTION
Accordingly, an object of the present invention is to provide a novel polypeptide having cold-stable PPDK activity, a cloned DNA encoding the same and a recombinant vector containing the DNA, as means for giving cold-stability to plants. Another object of the present invention is to provide a plant transformed with the above-mentioned DNA according to the present invention.
The present inventors intensively studied to succeed in cloning the complete PPDK gene of Flaveria brownii, determining the nucleotide sequence of the gene and the amino acid sequence encoded thereby, and in identifying the region in the PPDK gene, which gives cold-stability, thereby completing the present invention.
That is, the present invention provides a polypeptide having cold-stable PPDK activity, which has an amino acid sequence that is the same as the amino acid sequence of 1/6 region of the entire region from the C-terminal of the following polypeptide (1) or (2), except that at least one amino acid residues of said 1/6 region are substituted with other amino acid residues: Sequence Listing; and less than 50% with the amino acid sequence mentioned in (1), said polypeptide having cold-stable PPDK activity.
The present invention also provides a cloned DNA encoding the polypeptide having cold-stable PPDK activity according to the present invention. The present invention further provides a recombinant vector containing the DNA according to the present invention, which can express in a host a polypeptide having cold-stable PPDK activity. The present invention still further provides a plant which is transformed with the DNA according to the present invention.
By the present invention, a gene encoding PPDK having cold-stability was cloned and sequenced. Further, the region in the gene, which gives cold-stability was also identified. Therefore, by transforming a pla
REFERENCES:
Shimamoto et al, Nature, vol. 338, pp. 274-276 (Mar. 16, 1989).
Tanaka et al, Nucleic Acids Research, vol. 18, No. 23, pp. 6767-6770 (1990).
Baba et al, Plant Cell Physiol., 27(3), pp. 463-471 (1986).
Ueki et al, Plant Cell Physiol., 36(5), pp. 903-914 (1995).
Burnell, Plant Cell Physiol., 31(2), pp. 295-297 (1990).
Matsuoka et al. (1988) Primary Structure of Maize Pyruvate, Orthophosphate Dikinase as Deduced from cDNA Sequence, J. Biol. Chem. 263 (23): 11080-11083, Aug. 15, 1988.
Rosche et al. (1990) Primary structure of pyruvate, orthophosphate dikinase in the dicotyledonous C4 plant Flaveria trinervia, FEBS Letters 273 (1,2): 116-121, Oct. 1990.
Pocalyko et al. (1990) Analysis of Sequence Homologies in Plant and Bacterial Pyruvate Phosphate Dikinase, Enzyme I of the Bacterial . . . , Biochem. 29: 10757-10765, Nov. 1990.
Rosche et al. (1994) Primary structure of the photosynthetic pyruvate orthophosphate dikinase of the C3 plant Flaveria pringlei and . . . , Plant Molecular Biology 26: 763-769, Oct. 1994.
Rudinger, J. (1976) Characteristics of the amino acids as components of a peptide hormone sequence, In Peptide Hormones, Ed. J. A. Parsons, University Park Press, pp. 1-7, Jun. 1976.
Burnell James Nigel
Ohta Shozo
Usami Satoru
Japan Tobacco Inc.
Stole Einar
Wax Robert A.
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