Abstract
We isolated a complementary DNA sequence for the enzyme sucrose phosphate synthase (SPS) from maize utilizing a limited amino acid sequence. The 3509-bp cDNA encodes a 1068-amino acid polypeptide. The identity of the cDNA was confirmed by the ability of the cloned sequence to direct sucrose phosphate synthesis in Escherichia coli. Because no plant-specific factors were necessary for enzymatic activity, we can conclude that SPS enzyme activity is conferred by a single gene product. Sequence comparisons showed that SPS is distantly related to the enzyme sucrose synthase. When expressed from a ribulose bisphosphate carboxylase small subunit promoter in transgenic tomatoes, total SPS activity was boosted up to sixfold in leaves and appeared to be physiologically uncoupled from the tomato regulation mechanism. The elevated SPS activity caused a reduction of starch and increase of sucrose in the tomato leaves. This result clearly demonstrates that SPS is involved in the regulation of carbon partitioning in the leaves.
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- Bergman T., Jörnvall H. Electroblotting of individual polypeptides from SDS/polyacrylamide gels for direct sequence analysis. Eur J Biochem. 1987 Nov 16;169(1):9–12. doi: 10.1111/j.1432-1033.1987.tb13573.x. [DOI] [PubMed] [Google Scholar]
- Chourey P. S., Nelson O. E. The enzymatic deficiency conditioned by the shrunken-1 mutations in maize. Biochem Genet. 1976 Dec;14(11-12):1041–1055. doi: 10.1007/BF00485135. [DOI] [PubMed] [Google Scholar]
- Dickinson C. D., Altabella T., Chrispeels M. J. Slow-growth phenotype of transgenic tomato expressing apoplastic invertase. Plant Physiol. 1991 Feb;95(2):420–425. doi: 10.1104/pp.95.2.420. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Flügge U. I., Fischer K., Gross A., Sebald W., Lottspeich F., Eckerskorn C. The triose phosphate-3-phosphoglycerate-phosphate translocator from spinach chloroplasts: nucleotide sequence of a full-length cDNA clone and import of the in vitro synthesized precursor protein into chloroplasts. EMBO J. 1989 Jan;8(1):39–46. doi: 10.1002/j.1460-2075.1989.tb03346.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
- Kalt-Torres W., Huber S. C. Diurnal Changes in Maize Leaf Photosynthesis : III. Leaf Elongation Rate in Relation to Carbohydrates and Activities of Sucrose Metabolizing Enzymes in Elongating Leaf Tissue. Plant Physiol. 1987 Feb;83(2):294–298. doi: 10.1104/pp.83.2.294. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krieg P. A., Melton D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 1984 Sep 25;12(18):7057–7070. doi: 10.1093/nar/12.18.7057. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maraña C., García-Olmedo F., Carbonero P. Linked sucrose synthase genes in group-7 chromosomes in hexaploid wheat (Triticum aestivum L.). Gene. 1988 Mar 31;63(2):253–260. doi: 10.1016/0378-1119(88)90529-x. [DOI] [PubMed] [Google Scholar]
- McBride K. E., Summerfelt K. R. Improved binary vectors for Agrobacterium-mediated plant transformation. Plant Mol Biol. 1990 Feb;14(2):269–276. doi: 10.1007/BF00018567. [DOI] [PubMed] [Google Scholar]
- Mogen B. D., MacDonald M. H., Graybosch R., Hunt A. G. Upstream sequences other than AAUAAA are required for efficient messenger RNA 3'-end formation in plants. Plant Cell. 1990 Dec;2(12):1261–1272. doi: 10.1105/tpc.2.12.1261. [DOI] [PMC free article] [PubMed] [Google Scholar]
- O'Neal J. K., Pokalsky A. R., Kiehne K. L., Shewmaker C. K. Isolation of tobacco SSU genes: characterization of a transcriptionally active pseudogene. Nucleic Acids Res. 1987 Nov 11;15(21):8661–8677. doi: 10.1093/nar/15.21.8661. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salanoubat M., Belliard G. Molecular cloning and sequencing of sucrose synthase cDNA from potato (Solanum tuberosum L.): preliminary characterization of sucrose synthase mRNA distribution. Gene. 1987;60(1):47–56. doi: 10.1016/0378-1119(87)90212-5. [DOI] [PubMed] [Google Scholar]
- Stalker D. M., McBride K. E., Malyj L. D. Herbicide resistance in transgenic plants expressing a bacterial detoxification gene. Science. 1988 Oct 21;242(4877):419–423. doi: 10.1126/science.242.4877.419. [DOI] [PubMed] [Google Scholar]
- Thummler F., Verma D. P. Nodulin-100 of soybean is the subunit of sucrose synthase regulated by the availability of free heme in nodules. J Biol Chem. 1987 Oct 25;262(30):14730–14736. [PubMed] [Google Scholar]
- Van Handel E. Direct microdetermination of sucrose. Anal Biochem. 1968 Feb;22(2):280–283. doi: 10.1016/0003-2697(68)90317-5. [DOI] [PubMed] [Google Scholar]
- Werr W., Frommer W. B., Maas C., Starlinger P. Structure of the sucrose synthase gene on chromosome 9 of Zea mays L. EMBO J. 1985 Jun;4(6):1373–1380. doi: 10.1002/j.1460-2075.1985.tb03789.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- von Schaewen A., Stitt M., Schmidt R., Sonnewald U., Willmitzer L. Expression of a yeast-derived invertase in the cell wall of tobacco and Arabidopsis plants leads to accumulation of carbohydrate and inhibition of photosynthesis and strongly influences growth and phenotype of transgenic tobacco plants. EMBO J. 1990 Oct;9(10):3033–3044. doi: 10.1002/j.1460-2075.1990.tb07499.x. [DOI] [PMC free article] [PubMed] [Google Scholar]