Characterization of cDNAs encoding two isoforms of granule-bound starch synthase which show differential expression in developing storage organs of pea and potato

Dry, Ian B.; Smith, Alison M.; Edwards, Anne; Bhattacharyya, Madan K.; Dunn, P. P. J.; Martin, Cathie

doi:10.1111/j.1365-313x.1992.00193.x

Cited by 47 publications

(106 citation statements)

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“…The expression of GB S S in cassava was organspecific and correlated with starch deposition as has been reported in pea and potato [9,44]. Cassava tubers produced variable but high levels of mRNA at different stages of their development ensuring a continuous production and accumulation of starch (Fig.…”

Section: Isolation and Characterization Of The Gbss Cdna And Proteinmentioning

confidence: 80%

“…The GBSS (starch granule-bound ADP (UDP) glucose: e-1,4-D-glucan 4-e-glucosyl transferase, EC 2.4.1.21), also known as waxy protein, is a protein with a molecular mass of ca. 60 kDa in potato [49] and pea [9], 58 or 60 kDa in maize [40], 60 kDa in rice [38] and 68 kDa in grain amaranth [20]. Although GBS S catalyses in vitro the elongation of both amylose and amylopectin [32], mutations affecting the GBSS gene in several plant species resulted only in loss of amylose, while the total amount of starch remained more or less the same [39].…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, antisense GBSS RNA can suppress the 60 kDa protein in potato resulting in the absence of amylose from starch [47]. In pea, a protein of 77 kDa which can be solubilized from starch granules and which is antigenically only weakly related to the 60 kDa protein of potato, has been proposed as a second isoform of GB S S (GBSS II) [9]. However, its role in amylose synthesis is not clear.…”

Section: Introductionmentioning

confidence: 99%

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Isolation and characterization of a cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato

1993

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A tuber-specific cDNA library of cassava (Manihot esculenta Crantz) was constructed and a full-length cDNA for granule-bound starch synthase (GBSS, also known as waxy protein), the enzyme responsible for the synthesis of amylose in reserve starch, was cloned. Sequencing of the cloned cDNA showed that it has 74% identity with potato GBSS and 60-72% identity with GBSS from other plant species. The cDNA encodes a 608 amino acid protein of which 78 amino acids form a chloroplast/amyloplast transit peptide of 8.37 kDa. The mature protein has a predicted molecular mass of 58.61 kDa (530 amino acids). Comparison of the GBSS proteins of various plant species and glycogen synthase of bacteria showed extensive identity among the mature form of plant GBSS proteins, in which the monocots and dicots form two separate branches in the evolutionary tree. From analysis of the genomic DNA of allotetraploid cassava, it is shown that GBSS is a low-copy-number gene. GBSS transcript is synthesized in a number of different organs, but most abundantly in tubers. Potato plants were transformed with the cassava GBSS cDNA in antisense orientation fused between the potato GBSS promoter and the nopaline synthase terminator. The expression of the endogenous GBSS gene in these transgenic potato plants was partially or completely inhibited. Complete inhibition of GBSS activity by the cassava antisense gene resulted in absence of GBSS protein and amylose giving rise to almost complete amylose-free potato starch. This shows that also heterologous genes can be used to achieve antisense effects in other plant species.

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Section: Isolation and Characterization Of The Gbss Cdna And Proteinmentioning

confidence: 80%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Isolation and characterization of a cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato

1993

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“…These proteins have been termed GBSS1 and GBSSII respectively [5], and the authors propose that they represent functionally distinct isoforms of starch synthase which make different contributions to starch synthesis.…”

Section: Introductionmentioning

confidence: 99%

“…These include cDNA clones from pea [5], potato [40], barley [31] and wheat [4] and genomic clones from potato [32,40], rice [43], barley [31] and wheat (C.C. Ainsworth, unpublished).…”

Section: Introductionmentioning

confidence: 99%

Expression, organisation and structure of the genes encoding the waxy protein (granule-bound starch synthase) in wheat

1993

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A full-length cDNA clone representing the waxy protein (GBSSI) isolated from a hexaploid wheat developing grain cDNA library has been used to characterise the organisation and expression of the waxy genes in wheat. The genes are organised as a triplicate set of single copy homeoloci on chromosome arms 4AL, 7AS and 7DS. The genes are active throughout grain filling where the main 2.3 kb transcript accumulates to high levels. The 2.3 kb transcript is not expressed in leaves where the presence of a related, but less homologous, transcript of 1.6 kb suggests that a different set of genes operates. Gel analysis and purification of the waxy protein isolated from starch granules, followed by N-terminal amino acid sequencing in conjunction with data from hybrid select translation experiments and sequence analysis of the cDNA, shows that the mature protein has a molecular weight of 60kDa (615 amino acids) and that the preprotein includes a chloroplast/amyloplast transit peptide of 7kDa (75 amino acids). Analysis of the derived amino acid sequence and alignment with five other plant waxy proteins shows that they exhibit substantial homology. The wheat protein differs from all others in that it contains an 11 amino acid insertion towards the N-terminus. The protein contains the conserved motif KTGGL found in other waxy proteins and which has been implicated as the active site in glycogen synthase.

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A 38 kb segment containing thecdc2 gene from the left arm of fission yeast chromosome II: sequence analysis and characterization of the genomic DNA and cDNAs encoded on the segment

Machida¹,

Yamazaki

Kunihiro³

et al. 2000

Yeast

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A genomic 38 kbp segment on the c1750 cosmid clone containing the cdc2 gene, located in the left arm of chromosome II from Schizosaccharomyces pombe, was sequenced. The segment was found to have ®ve previously known genes, pht1, cdc2, his3, act1 and mei4. Among 11 coding sequences (CDSs) predicted by the gene ®nding software INTRON.PLOT., four CDSs, pi007, pi010, pi014 and pi016, had considerable similarity to 40S ribosomal protein, glycosyltransferase, cdc2-related protein kinase and a-1,2-mannosyltransferase, respectively. Another unusually huge open reading frame (ORF) (pi011), consisting of 2233 amino acids, existed, having signi®cant homology to a-amylase, granule-bound glycogen synthase and the Sz. pombe YS 1110 clone product at the N-terminal, middle and C-terminal regions, respectively. All the predicted 11 CDSs were experimentally analysed by RACE PCR. The sequencing of the RACE products revealed that there were two small overlaps at the 3k untranslated regions (UTRs) between pi004 and pi005 (17 bp) and between pi007 and pi008 (2 bp). The distances between 5k end of the 5kUTR and the putative translation initiation codon varied from 10 to 302 nucleotides (nt) among the nine CDSs successfully analysed by 5k-RACE. The expression level of each CDS on this clone was determined. Among the 16 genes on this clone, the previously determined genes, pht1, cdc2, his3 and act1, were found to be most highly expressed. Finally, cDNAs of all the newly identi®ed genes were detected by RACE, proving the actual expression of these genes. The nucleotide sequence has been submitted to the EMBL database under Accession No. AB004534.

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Characterization of cDNAs encoding two isoforms of granule-bound starch synthase which show differential expression in developing storage organs of pea and potato

Cited by 47 publications

References 40 publications

Isolation and characterization of a cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato

Isolation and characterization of a cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato

Expression, organisation and structure of the genes encoding the waxy protein (granule-bound starch synthase) in wheat

A 38 kb segment containing thecdc2 gene from the left arm of fission yeast chromosome II: sequence analysis and characterization of the genomic DNA and cDNAs encoded on the segment

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