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  • 1
    ISSN: 1573-5079
    Keywords: PEP carboxylase ; C3 ; C4
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this report, the effects of light on the activity and allosteric properties of phosphoenolpyruvate (PEP) carboxylase were examined in newly matured leaves of several C3 and C4 species. Illumination of previously darkened leaves increased the enzyme activity 1.1 to 1.3 fold in C3 species and 1.4 to 2.3 fold in C4 species, when assayed under suboptimal conditions (pH 7) without allosteric effectors. The sensitivities of PEP carboxylase to the allosteric effectors malate and glucose-6-phosphate were markedly different between C3 and C4 species. In the presence of 5 mM malate, the activity of the enzyme extracted from illuminated leaves was 3 to 10 fold higher than that from darkened leaves in C4 species due to reduced malate inhibition of the enzyme from illuminated leaves, whereas it increased only slightly in C3 species. The Ki(malate) for the enzyme increased about 3 fold by illumination in C4 species, but increased only slightly in C3 species. Also, the addition of the positive effector glucose-6-phosphate provided much greater protection against malate inhibition of the enzyme from C4 species than C3 species. Feeding nitrate to excised leaves of nitrogen deficient plants enhanced the degree of light activation of PEP carboxylase in the C4 species maize, but had little or no effect in the C3 species wheat. These results suggest that post-translational modification by light affects the activity and allosteric properties of PEP carboxylase to a much greater extend in C4 than in C3 species.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5028
    Keywords: C4 photosynthesis ; maize ; phosphoenolpyruvate carboxylase ; transgenic plant ; transcription ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract C4-type phosphenolpyruvate carboxylase (C4PEPC) acts as a primary carbon assimilatory enzyme in the C4 photosynthetic pathway. The maize C4PEPC gene (C4Ppc1) is specifically expressed in mesophyll cells (MC) of light-grown leaves, but the molecular mechanism responsible for its cell type-specific expression has not been characterized. In this study, we introduced a chimeric maize C4Ppc1 5′-flanking region/β-glucuronidase (GUS) gene into maize plants by Agrobacterium-mediated transformation. Activity assay and histochemical staining showed that GUS is almost exclusively localized in leaf MC of transgenic maize plants. This observation suggests that the introduced 5′ region of maize C4Ppc1 contains the necessary cis element(s) for its specific expression in MC. Next, we investigated whether the 5′ region of the maize gene interacts with nuclear proteins in a cell type-specific manner. By gel shift assays with nuclear extracts prepared from MC or bundle sheath cells (BSC), cell type-specific DNA-protein interactions were detected: nuclear factors PEPIb and PEPIc are specific to MC whereas PEPIa and PEPIIa are specific to BSC. Light alters the binding activity of these factors. These interactions were not detected in the assay with nuclear extract prepared from root, or competed out by oligonucleotides corresponding to the binding sites for the maize nuclear protein, PEP-I, which is known to bind specifically to the promoter region of C4Ppc1. The results suggest that novel cell type-specific positive and negative nuclear factors bind to the maize C4Ppc1 5′-flanking region and regulate its differential transcription in MC in a light-dependent manner.
    Type of Medium: Electronic Resource
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