Abstract
l-myo-Inositol 1-phosphate synthase (EC 5.5.1.4; MIPS) catalyzes conversion of glucose 6-phosphate to l-myo-inositol 1-phosphate, the first and the rate-limiting step in the production of inositol, and has been reported from evolutionarily diverse organisms. Two forms of the enzyme have been characterized from higher plants, viz. cytosolic and chloroplastic, and the presence of MIPS has been earlier reported from the cyanobacteria (e.g. Spirulina sp.), the presumed chloroplast progenitors. The present study demonstrates possible multiple forms of MIPS and identifies the gene for one of them in the cyanobacterium Synechocystis sp. PCC 6803. Following detection of at least two immunologically cross-reactive MIPS forms, we have been able to identify from the fully sequenced Synechocystis genome an as yet unassigned open reading frame (ORF), sll1722, coding for the approx. 50-kDa MIPS protein, by using biochemical, molecular and bioinformatics tools. The DNA fragment corresponding to sll1722 was PCR-amplified and functional identity of the gene was confirmed by a complementation assay in Saccharomyces cerevisiae mutants containing a disrupted INO1 gene for the yeast MIPS. The sll1722 PCR product was cloned in Escherichia coli expression vector pET20b and the isopropyl β-d-thiogalactopyranoside (IPTG)-induced overexpressed protein product was characterized following complete purification. Comparison of the sll1722 sequences with other MIPS sequences and its phylogenetic analysis revealed that the Synechocystis MIPS gene is quite divergent from the others.
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Abbreviations
- CBB :
-
Coomassie Brilliant Blue
- EST :
-
Expressed sequence tag
- G6P :
-
d-Glucose 6-phosphate
- IPTG:
-
Isopropyl β-d-thiogalactopyranoside
- MIPS :
-
l–myo-Inositol 1-phosphate synthase
- ORF:
-
Open reading frame
References
Adhikari J, Majumder AL, Bhaduri TJ, DasGupta S, Majumder AL (1987) Chloroplast as a locale of l-myo-inositol 1-phosphate synthase. Plant Physiol 85:611–614
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
Bachhawat N, Mande SC (1999) Identification of the INO1 gene of Mycobacterium tuberculosis H37Rv reveals a novel class of inositol 1-phosphate synthase enzyme. J Mol Biol 291:531–536
Bachhawat N, Mande SC (2000) Complex evolution of the inositol-1-phosphate synthase gene among archaea and eubacteria. Trends Genet 16:111–113
Barnett JEG, Brice RE, Corina DL (1970) A colorimetric determination of inositol monophosphates as an assay for d-glucose-6-phosphate-1l-myo-inositol 1-phosphate cyclase. Biochem J 119:183–186
Biswas BB, Ghosh B, Majumder AL (1984) Myo-inositol polyphosphates and their role in cellular metabolism—a proposed cycle involving glucose-6-phosphate and myo-inositol phosphates. In: Roodyn DB (ed) Subcellular biochemistry, vol 10. Plenum Press, London, pp 237–280
Bohnert HJ, Nelson DE, Jensen RG (1995) Adaptations to environmental stresses. Plant Cell 7:1099–1111
Bradford MM (1976) Quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254
Chen L, Zhou C, Yang H, Roberts MF (2000) Inositol-1-phosphate from Archaeoglobus fulgidus is a Class II aldolase. Biochemistry 39:12415–12423
Chen PS, Toribara TY, Warner H (1956) Microdetermination of phosphorus. Anal Chem 28:1756
DasGupta S, Adhikari J, Majumder AL (1984) l-myo-inositol-1-phosphate synthase from lower plant groups: partial purification and properties of the enzyme from Euglena gracilis. Physiol Plant 61:412–416
Donahue TF, Henry SA (1981) Myo-inositol 1-phosphate synthase: characteristics of the enzyme and identification of its structural gene in yeast. J Biol Chem 256:7077–7085
Gray MW (1991) Origin and evolution of plastid genomes and genes. In: Bogorad I, Vasil IK (eds) Molecular biology of plastids. Academic press, San Diego, pp 303–313
Gray MW (1992) The endosymbiotic hypothesis revisited. Int Rev Cytol 141:233–357
Hait NC, RayChaudhuri A, Das A, Bhattacharyya S, Majumder AL (2002) Processing and activation of chloroplast l-myo-inositol 1-phosphate synthase from Oryza sativa requires signals from both light and salt. Plant Sci 162:559–568
Imhoff V, Bourdu R (1973) Formation, d’ inositol par les chloroplasts isoles de pois. Phytochemistry 12:331–336
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
Jin X, Geiger JH (2003) Structures of NAD(+)- and NADH-bound 1-l–myo-inositol 1-phosphate synthase. Acta Crystallogr Ser D 59:1154–1164
Johnson MD, Henry SA (1989) Biosynthesis of inositol in yeast: primary structure of myo-inositol 1-phosphate synthase locus and functional characterization of its structural gene, the Ino1 locus. J Biol Chem 264:1274–1283
Kelley LA, MacCallum RM, Sternberg MJE (2000) Enhanced genome annotation using structural profiles in the program 3D-PSSM. J Mol Biol 299:499–520
Klig LS, Henry SA (1984) Isolation of INO1 gene on an autonomously replicating plasmid, the gene is fully regulated. Proc Natl Acad Sci USA 81:3816–3820
Laemmli UK (1970) Cleavages of structural proteins during assembly of the head of bacteriophage T4. Nature 227:680–685
Lohia A, Hait NC, Majumder AL (1999) l-myo-Inositol 1-phosphate synthase from Entamoeba histolytica. Mol Biochem Parasitol 98:67–79
Loewus FA, Murthy PPN (2000) Myo-inositol metabolism in Plants. Plant Sci 150:1–19
Maeda T, Eisenberg F Jr (1980) Purification, structure, and catalytic properties of l-myo-inositol-1-phosphate synthase from rat testis. J Biol Chem 255:8458–8464
Majumder AL, Duttagupta S, Goldwasser P, Donahue TF, Henry SA (1981) The mechanism of interallelic complementation at the INO1 locus in yeast: immunological analysis of mutants. Mol Gen Genet 184:347–354
Majumder AL, Johnson MD, Henry SA (1997) 1l-myo-Inositol-1–phosphate synthase. Biochim Biophys Acta 1348:245–256
Majumder AL, Chatterjee A, GhoshDastidar K, Majee M (2003) Diversification and evolution of l-myo-inositol 1-phosphate synthase. FEBS Lett 533:3–10
Mazur BJ, Rice D, Haselkorn R (1980) Identification of blue green algal nitrogen fixation genes by using heterologous DNA hybridization probes. Proc Natl Acad Sci USA 77:186–190
Mumburg D, Muller R, Funk M (1995) Yeast vectors for the controlled expression of heterologous proteins in different genetic backgrounds. Gene 156:119–122
Norman RA, McAlister MSB, Murray-Rust J, Movahedzadeh F, Stoker NG, McDonald NQ (2002) Crystal structure of inositol 1-phosphate synthase from Mycobacterium tuberculosis, a key enzyme in phosphatidylinositol synthesis. Structure 10:393–402
RayChaudhuri A, Majumder AL (1996) Salinity induced enhancement of l-myo-inositol 1-phosphate synthase in rice (Oryza sativa L.). Plant Cell Environ 19:1337–1342
RayChaudhuri A, Hait NC, DasGupta S, Bhaduri TJ, Deb R, Majumder AL (1997) l-myo-inositol 1-phosphate synthase from plant sources: characteristics of the chloroplastic and cytosolic enzymes. Plant Physiol 115:727–736
Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111:1–6
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Stein AJ, Geiger J H (2000) Structural studies of MIP synthase. Acta Crystallogr Ser D 56:348–350
Stein AJ, Geiger JH (2002) The crystal structure and mechanism of 1-l-myo-inositol-1-phosphate synthase. J Biol Chem 277:9484–9491
Acknowledgements
A.C. thanks the Council of Scientific and Industrial Research, Government of India for a Senior Research Fellowship, and the Bioinformatics Centre of Bose Institute for facilities. The work is supported by grants to A.L.M. from the Department of Biotechnology, Department of Science and Technology and Council of Scientific and Industrial Research, Government of India.
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Chatterjee, A., Majee, M., Ghosh, S. et al. sll1722, an unassigned open reading frame of Synechocystis PCC 6803, codes for l-myo-inositol 1-phosphate synthase. Planta 218, 989–998 (2004). https://doi.org/10.1007/s00425-003-1190-5
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DOI: https://doi.org/10.1007/s00425-003-1190-5