Isolation and expression analysis of cDNA clones encoding a small and a large subunit of ADP-glucose pyrophosphorylase from sugar beet

Müller‐Röber, Bernd; Nast, G.; Willmitzer, Lothar

doi:10.1007/bf00019190

Cited by 14 publications

(6 citation statements)

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“…RNA gel blot analysis of expression of the large subunit gene of sugar beet indicated that the large subunit and small subunit genes were strongly expressed in sink and source leaves (Müller-Röber et al 1995). A high level of mRNA accumulation for the large subunit gene was observed in the stem of sweet potato, but it was totally absent in the tuber and photosynthetic leaves (Harn et al 2000; Lee et al 1999).…”

Section: Discussionmentioning

confidence: 99%

Activity and expression of ADP-glucose pyrophosphorylase during rhizome formation in lotus (Nelumbo nucifera Gaertn.)

et al. 2016

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BackgroundLotus root is a traditional and popular aquatic vegetable in China. Starch is an important component of the rhizome and directly affects the quality of processed products. ADP -glucose pyrophosphorylase (AGPase) is a rate-limiting enzyme associated with starch biosynthesis in plants. Therefore, in the present study, AGPase activity and NnAGP expression during rhizome development of lotus were analyzed.ResultsAmong 15 cultivars analyzed, the contents of amylose and total starch in the rhizome were highest in ‘Mei Ren Hong’. ‘Su Zhou’ and ‘Zhen Zhu’ showed the lowest amylose, amylopectin and total starch contents. In the rhizome, activity of AGPase was highest at the middle swelling stage of development, and higher activity was observed in the ‘Hou ba’ leaf and terminational leaf at the same stage. Three AGPase genes, comprising two large subunit genes (NnAGPL1 and NnAGPL2) and one small subunit gene (NnAGPS), were isolated and identified. The deduced amino acid sequences showed 40.5 % similarity among the three genes. Full-length genomic DNA sequences of NnAGPL1, NnAGPL2, and NnAGPS were 4841, 11,346 and 4169 bp, respectively. Analysis of the temporal and spatial expression patterns revealed that the transcription levels of NnAGPL1 and NnAGPS were higher in the rhizome, followed by the ‘Hou ba’ leaf, whereas NnAGPL2 was significantly detected in the ‘Hou ba’ leaf and terminational leaf. The initial swelling stage of rhizome development was accompanied by the highest accumulation of mRNAs of NnAGPL1, whereas expression of NnAGPL2 was not detected during rhizome development. The transcript level of NnAGPS was highest at the initial swelling stage compared with the other rhizome developmental stages. Transcription of NnAGPL1, NnAGPL2, and NnAGPS was induced within 24 h after treatment with exogenous sucrose. The mRNA level of NnAGPL1 and NnAGPS was increased by exogenous ABA, whereas transcription of NnAGPL2 was not affected by ABA.ConclusionsThe three AGPase genes display marked differences in spatial and temporal expression patterns. Regulation of AGPase in relation to starch synthesis in lotus is indicated to be complex.Electronic supplementary materialThe online version of this article (doi:10.1186/s40529-016-0140-z) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Activity and expression of ADP-glucose pyrophosphorylase during rhizome formation in lotus (Nelumbo nucifera Gaertn.)

et al. 2016

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“…The only time that the Lys residue of the site s-I was not conserved was in a clone from sugar beet that had a deletion of 12 amino acids in that region. However, this clone has not been expressed to determine if it has any activity and it was used only as a probe in blot experiments (Muller-Rö ber et al, 1995). Another exception is a PCR product from Arabidopsis.…”

Section: Discussionmentioning

confidence: 99%

ADP-Glucose Pyrophosphorylase from Potato Tubers. Site-Directed Mutagenesis Studies of the Regulatory Sites1

Ballícora

Nesbitt

et al. 1998

Plant Physiology

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The apparent affinity for the inhibitor, phosphate, decreased greater than 400-fold. Mutation of Lys-441 to glutamic acid showed even larger effects. When Lys-417 and Lys-455 on the large subunit were mutated to alanine, the phosphate inhibition was not altered and the apparent affinity for the activator decreased only 9-and 3-fold, respectively. Mutations of these residues to glutamic acid only decreased the affinity for the activator 12-and 5-fold, respectively. No significant changes were observed on other kinetic constants for the substrates ADP-Glc, pyrophosphate, and Mg 2؉ . These data indicate that Lys-404 and Lys-441 on the small subunit are more important for the regulation of the ADP-Glc pyrophosphorylase than their homologous residues in the large subunit.

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“…The LS group can be divided into two subgroups corresponding to sink (sites of net sugar import like endosperm, roots, and young leaves) and source (sites of net sugar export like mature leaves) tissues, with the exception of the sugar beet large subunit that is expressed in sink and source leaves (27) and is included in the sink group of LS. Arabidopsis APL1 is considered to be the main isoform expressed in leaves (36).…”

Section: Fig 2 Activation By 3-pga Of the Recombinant Arabidopsis Amentioning

confidence: 99%

“…3, SS genes are expressed in both photosynthetic and nonphotosynthetic tissues, whereas LS genes are expressed preferentially in a tissue-specific manner (9,23,27). Arabidopsis ApL1 is mainly expressed in leaves (36) and, correspondingly, is included in the leaf group of APLs (Fig.…”

Section: Fig 3 Comparison Of Adp-glcmentioning

confidence: 99%

“…Different isoforms for ADP-Glc PPase subunits have been described, and many cDNAs and genomic DNAs encoding for them have been isolated from both monocot and dicot plants. The most frequent situation is the existence of one SS gene and several LS genes that are differentially expressed (3,9,(23)(24)(25)(26)(27)(28)(29). In Arabidopsis one SS gene (ApS1) and three LS genes (ApL1, ApL2, and ApL3) have been previously identified (30).…”

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The Different Large Subunit Isoforms of Arabidopsis thaliana ADP-glucose Pyrophosphorylase Confer Distinct Kinetic and Regulatory Properties to the Heterotetrameric Enzyme

Crevillén

Ballícora

Mérida

et al. 2003

Journal of Biological Chemistry

126

153

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ADP-glucose pyrophosphorylase catalyzes the first and limiting step in starch biosynthesis and is allosterically regulated by the levels of 3-phosphoglycerate and phosphate in plants. ADP-glucose pyrophosphorylases from plants are heterotetramers composed of two types of subunits (small and large). In this study, the six Arabidopsis thaliana genes coding for ADP-glucose pyrophosphorylase isoforms (two small and four large subunits) have been cloned and expressed in an Escherichia coli mutant deficient in ADP-glucose pyrophosphorylase activity. The co-expression of the small subunit APS1 with the different Arabidopsis large subunits (APL1, APL2, APL3, and APL4) resulted in heterotetramers with different regulatory and kinetic properties. Heterotetramers composed of APS1 and APL1 showed the highest sensitivity to the allosteric effectors as well as the highest apparent affinity for the substrates (glucose-1-phosphate and ATP), whereas heterotetramers formed by APS1 and APL2 showed the lower response to allosteric effectors and the lower affinity for the substrates. No activity was detected for the second gene coding for a small subunit isoform (APS2) annotated in the Arabidopsis genome. This lack of activity is possibly due to the absence of essential amino acids involved in catalysis and/or in the binding of glucose-1-phosphate and 3-phosphoglycerate. Kinetic and regulatory properties of the different heterotetramers, together with sequence analysis has allowed us to make a distinction between sink and source enzymes, because the combination of different large subunits would provide a high plasticity to ADP-glucose pyrophosphorylase activity and regulation. This is the first experimental data concerning the role that all the ADP-glucose pyrophosphorylase isoforms play in a single plant species. This phenomenon could have an important role in vivo, because different large subunits would confer distinct regulatory properties to ADP-glucose pyrophosphorylase according to the necessities for starch synthesis in a given tissue.

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Isolation and expression analysis of cDNA clones encoding a small and a large subunit of ADP-glucose pyrophosphorylase from sugar beet

Cited by 14 publications

References 40 publications

Activity and expression of ADP-glucose pyrophosphorylase during rhizome formation in lotus (Nelumbo nucifera Gaertn.)

Activity and expression of ADP-glucose pyrophosphorylase during rhizome formation in lotus (Nelumbo nucifera Gaertn.)

ADP-Glucose Pyrophosphorylase from Potato Tubers. Site-Directed Mutagenesis Studies of the Regulatory Sites1

The Different Large Subunit Isoforms of Arabidopsis thaliana ADP-glucose Pyrophosphorylase Confer Distinct Kinetic and Regulatory Properties to the Heterotetrameric Enzyme

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