Isolation, characterization, and expression analysis of starch synthase IIa cDNA from wheat (Triticum aestivum L.)

Genome ◽  
2000 ◽  
Vol 43 (5) ◽  
pp. 768-775 ◽  
Author(s):  
Ming Gao ◽  
Ravindra N Chibbar
Keyword(s):  
Recombinant Protein ◽  
Polyclonal Antibody ◽  
Sequence Similarity ◽  
Triticum Aestivum L ◽  
Biochemical Properties ◽  
Starch Biosynthesis ◽  
Starch Synthase ◽  
Cdna Clones ◽  
Wheat Endosperm ◽  
Starch Synthases

We characterized three near-full-length putative homoeologous cDNA (Ss2a-1, Ss2a-2, and Ss2a-3) in wheat endosperm most similar to the maize zSSIIa. Polypeptide sequences deduced from three Ss2a cDNA clones share a 95% overall sequence similarity, and may thus have similar biochemical properties and may make identical contributions to starch biosynthesis in wheat endosperm. The accumulation of RNA transcripts corresponding to three Ss2a genes in developing endosperm varies among three cultivars studied, but usually peaks in young endosperm at about 10 days post anthesis (DPA). The polyclonal antibody for the SSIIa-1 recombinant protein strongly reacted to three previously identified granule-bound starch synthases of 100 to 115 kDa. The polyclonal antibody for the granule-bound starch synthases strongly reacted to the SSIIa-1 recombinant protein. Sequences of the N-terminal and an internal peptide of these three granule-bound starch synthases match well with those of three predicted mature SSIIa polypeptides. These granule-bound starch synthases may therefore be SSIIa polypeptides. The antibodies also recognized a group of three polypeptides with the same gel mobility as the three granule-bound starch synthases, a polypeptide of 90 kDa, and a group of three polypeptides of about 80 to 82 kDa. Thus, the wheat SSIIa may exist in several functional forms in the stroma of amyloplasts.Key words: starch granule, granule-bound proteins, soluble starch synthase, homoeologous isoforms, starch biosynthesis.

scholarly journals Biosynthesis of waxy starch – a review

2017 ◽  
Vol 63 (No. 8) ◽  
pp. 335-341 ◽  
Author(s):  
Šárka Evžen ◽  
Dvořáček Václav
Keyword(s):  
Plant Breeding ◽  
Metabolic Pathways ◽  
Special Form ◽  
Starch Biosynthesis ◽  
Starch Synthase ◽  
Conversion Process ◽  
Starch Synthases ◽  
Competent Cells ◽  
Waxy Starch ◽  
Starch Conversion

Starch comprises nearly linear amylose and branched amylopectin, whilst waxy starches are a special form, containing almost exclusively amylopectin. Modern techniques in plant breeding together with new data from starch biosynthesis research have enabled new food and non-food uses of waxy starches. This paper describes the basic ways of glucose conversion to waxy starch in plants. The recent evidence of ADP-Glc accumulation in cytosol of photosynthetically competent cells proposes a more complex pathway of starch biosynthesis based on a tight interconnection of sucrose and starch metabolic pathways. Also many studies indicate the existence of different pathways for the sucrose-starch conversion process in heterotrophic organs of dicotyledonous and monocotyledonous plants. At least six classes of starch synthases (SS) have been recognised in plants including soluble SS1, SS2, SS3, SS4, SS5, and granule bound SS (GBSS), required for the synthesis of short and long chains of amylopectin, till now. As to amylose (not-present in waxy starches), GBSS is the only starch synthase isoform encoded by the waxy genes situated at independent loci.

scholarly journals The Pichia pastoris PER6 gene product is a peroxisomal integral membrane protein essential for peroxisome biogenesis and has sequence similarity to the Zellweger syndrome protein PAF-1.

1996 ◽  
Vol 16 (5) ◽  
pp. 2527-2536 ◽  
Author(s):  
H R Waterham ◽  
Y de Vries ◽  
K A Russel ◽  
W Xie ◽  
M Veenhuis ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Membrane Protein ◽  
Sequence Similarity ◽  
Zellweger Syndrome ◽  
Methylotrophic Yeast ◽  
Integral Membrane Protein ◽  
Biochemical Properties ◽  
Podospora Anserina ◽  
Peroxisome Biogenesis ◽  
Membrane Spanning

We report the cloning of PER6, a gene essential for peroxisome biogenesis in the methylotrophic yeast Pichia pastoris. The PER6 sequence predicts that its product Per6p is a 52-kDa polypeptide with the cysteine-rich C3HC4 motif. Per6p has significant overall sequence similarity with the human peroxisome assembly factor PAF-1, a protein that is defective in certain patients suffering from the peroxisomal disorder Zellweger syndrome, and with car1, a protein required for peroxisome biogenesis and caryogamy in the filamentous fungus Podospora anserina. In addition, the C3HC4 motif and two of the three membrane-spanning segments predicted for Per6p align with the C3HC4 motifs and the two membrane-spanning segments predicted for PAF-1 and car1. Like PAF-1, Per6p is a peroxisomal integral membrane protein. In methanol- or oleic acid-induced cells of per6 mutants, morphologically recognizable peroxisomes are absent. Instead, peroxisomal remnants are observed. In addition, peroxisomal matrix proteins are synthesized but located in the cytosol. The similarities between Per6p and PAF-1 in amino acid sequence and biochemical properties, and between mutants defective in their respective genes, suggest that Per6p is the putative yeast homolog of PAF-1.

scholarly journals Suppression of starch synthase I expression affects the granule morphology and granule size and fine structure of starch in wheat endosperm

2014 ◽  
Vol 65 (8) ◽  
pp. 2189-2201 ◽  
Author(s):  
S. J. McMaugh ◽  
J. L. Thistleton ◽  
E. Anschaw ◽  
J. Luo ◽  
C. Konik-Rose ◽  
...  
Keyword(s):  
Fine Structure ◽  
Granule Size ◽  
Starch Synthase ◽  
Wheat Endosperm ◽  
Granule Morphology

Multiple effects of the starch synthase II mutation in developing wheat endosperm

2007 ◽  
Vol 34 (5) ◽  
pp. 431 ◽  
Author(s):  
Behjat Kosar-Hashemi ◽  
Zhongyi Li ◽  
Oscar Larroque ◽  
Ahmed Regina ◽  
Makoto Yamamori ◽  
...  
Keyword(s):  
Triticum Aestivum L ◽  
Endosperm Development ◽  
Starch Synthase ◽  
Starch Granules ◽  
Branching Enzyme ◽  
Wild Type ◽  
Drastic Reduction ◽  
Branching Patterns ◽  
Soluble Phase ◽  
Mutant Lines

A line of wheat (Triticum aestivum L.), sgp-1, that does not express starch synthase II (SSII, also known as SGP-1) has previously been reported. In this study, F1 derived doubled haploid lines with homozygous wild type or mutant alleles for SGP-1 genes were identified from a cross between the original mutant and a wild type Australian cultivar. Analysis of the starch granules showed that in the mutant lines they are markedly distorted from 15 days postanthesis during grain development. Starch branching patterns showed an increase in the proportion of short chains (DP 6–10) at an earlier stage, but this increase became much more pronounced at 15 days postanthesis and persisted until maturity. There was also a consistent and drastic reduction throughout seed development in the relative amounts of starch branching enzyme II (SBEII, comprising SBEIIa and SBEIIb) and starch synthase I (SSI) bound to the starch granules. In the soluble phase, however, there was relatively little change in the amount of SBEIIb, SBEIIa or SSI protein. Therefore loss of SSII specifically leads to the loss of SBEIIb, SBEIIa and SSI protein in the granule-bound phase and the effect of this mutation is clearly manifest from the mid-stage of endosperm development in wheat.

scholarly journals cDNA cloning of human and rat brain myo-inositol monophosphatase. Expression and characterization of the human recombinant enzyme

1992 ◽  
Vol 284 (3) ◽  
pp. 749-754 ◽  
Author(s):  
G McAllister ◽  
P Whiting ◽  
E A Hammond ◽  
M R Knowles ◽  
J R Atack ◽  
...  
Keyword(s):  
Rat Brain ◽  
Cell Signalling ◽  
Biochemical Properties ◽  
Recombinant Enzyme ◽  
Cdna Clones ◽  
Coding Region ◽  
Inositol Monophosphatase ◽  
Human Enzyme ◽  
Key Enzyme

Inositol monophosphatase (EC 3.1.3.25) is a key enzyme in the phosphoinositide cell-signalling system. Its role is to provide inositol required for the resynthesis of phosphatidylinositol and polyphosphoinositides. It is the probable pharmacological target for lithium action in brain. Using probes derived from the bovine inositol monophosphatase cDNA we have isolated cDNA clones encoding the human and rat brain enzymes. The enzyme is highly conserved in all three species (79% identical). The coding region of the human cDNA was inserted into a bacterial expression vector. The expressed recombinant enzyme was purified and its biochemical properties examined. The human enzyme is very similar to the bovine enzyme.

Structure and expression of histone H3.3 genes in Drosophila melanogaster and Drosophila hydei

Genome ◽  
1995 ◽  
Vol 38 (3) ◽  
pp. 586-600 ◽  
Author(s):  
Anna S. Akhmanova ◽  
Petra C. T. Bindels ◽  
Jie Xu ◽  
Koos Miedema ◽  
Hannie Kremer ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Sequence Similarity ◽  
Expression Patterns ◽  
Alternative Polyadenylation ◽  
Cdna Clones ◽  
Drosophila Hydei ◽  
Histone H3.3 ◽  
Somatic Tissues ◽  
Polyadenylation Sites ◽  
Limited Sequence

We demonstrate that in Drosophila melanogaster the histone H3.3 replacement variant is encoded by two genes, H3.3A and H3.3B. We have isolated cDNA clones for H3.3A and cDNA and genomic clones for H3.3B. The genes encode exactly the same protein but are widely divergent in their untranslated regions (UTR). Both genes are expressed in embryos and adults; they are expressed in the gonads as well as in somatic tissues of the flies. However, only one of them, H3.3A, shows strong testes expression. The 3′ UTR of the H3.3A gene is relatively short (~250 nucleotides (nt)). H3.3B transcripts can be processed at several polyadenylation sites, the longest with a 3′ UTR of more than 1500 nt. The 3′ processing sites, preferentially used in the gonads and somatic tissues, are different. We have also isolated the Drosophila hydei homologues of the two H3.3 genes. They are quite similar to the D. melanogaster genes in their expression patterns. However, in contrast to their vertebrate counterparts, which are highly conserved in their noncoding regions, the Drosophila genes display only limited sequence similarity in these regions.Key words: H3.3 histone variant, Drosophila, sequence comparison, alternative polyadenylation, testis expression.

scholarly journals Taxonomic Characterization, and Secondary Metabolite Analysis of Streptomyces triticiradicis sp. nov.: A Novel Actinomycete with Antifungal Activity

2020 ◽  
Vol 8 (1) ◽  
pp. 77 ◽  
Author(s):  
Zhiyin Yu ◽  
Chuanyu Han ◽  
Bing Yu ◽  
Junwei Zhao ◽  
Yijun Yan ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
16S Rrna ◽  
Sequence Similarity ◽  
Triticum Aestivum L ◽  
Biosynthetic Gene Cluster ◽  
Rrna Gene ◽  
Phenotypic Properties ◽  
Genus Streptomyces ◽  
Antagonistic Microorganisms ◽  
Taxonomic Characterization

The rhizosphere, an important battleground between beneficial microbes and pathogens, is usually considered to be a good source for isolation of antagonistic microorganisms. In this study, a novel actinobacteria with broad-spectrum antifungal activity, designated strain NEAU-H2T, was isolated from the rhizosphere soil of wheat (Triticum aestivum L.). 16S rRNA gene sequence similarity studies showed that strain NEAU-H2T belonged to the genus Streptomyces, with high sequence similarities to Streptomyces rhizosphaerihabitans NBRC 109807T (98.8%), Streptomyces populi A249T (98.6%), and Streptomyces siamensis NBRC 108799T (98.6%). Phylogenetic analysis based on 16S rRNA, atpD, gyrB, recA, rpoB, and trpB gene sequences showed that the strain formed a stable clade with S. populi A249T. Morphological and chemotaxonomic characteristics of the strain coincided with members of the genus Streptomyces. A combination of DNA–DNA hybridization results and phenotypic properties indicated that the strain could be distinguished from the abovementioned strains. Thus, strain NEAU-H2T belongs to a novel species in the genus Streptomyces, for which the name Streptomyces triticiradicis sp. nov. is proposed. In addition, the metabolites isolated from cultures of strain NEAU-H2T were characterized by nuclear magnetic resonance (NMR) and mass spectrometry (MS) analyses. One new compound and three known congeners were isolated. Further, genome analysis revealed that the strain harbored diverse biosynthetic potential, and one cluster showing 63% similarity to natamycin biosynthetic gene cluster may contribute to the antifungal activity. The type strain is NEAU-H2T (= CCTCC AA 2018031T = DSM 109825T).

scholarly journals PfPK6, a novel cyclin-dependent kinase/mitogen-activated protein kinase-related protein kinase from Plasmodium falciparum

2000 ◽  
Vol 347 (1) ◽  
pp. 255-263 ◽  
Author(s):  
Valerie BRACCHI-RICARD ◽  
Sailen BARIK ◽  
Cherie DELVECCHIO ◽  
Christian DOERIG ◽  
Ratna CHAKRABARTI ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Protein Kinase ◽  
Catalytic Domain ◽  
Sequence Similarity ◽  
Small Subunit ◽  
Mitogen Activated Protein Kinase ◽  
Cdna Clones ◽  
Cyclin Dependent Kinase ◽  
Mitogen Activated Protein

We have isolated a novel protein kinase cDNA, PfPK6, by differential display RT-PCR (DDRT-PCR) of mRNA obtained from different asexual erythrocytic stages of Plasmodium falciparum, which shows sequence similarity to both cyclin-dependent kinase (CDK) and mitogen-activated protein kinase (MAPK) family members. The 915 bp open reading frame (ORF) is interrupted by seven introns and encodes a 305-residue polypeptide with a predicted molecular mass of 35848 Da. Several cDNA clones with some of the intron sequences were isolated, indicating alternate or defective splicing of PfPK6 transcripts because the gene seems to be a single copy located on chromosome 13. The similarity of the catalytic domain of PfPK6 to those of CDK2 and MAPK is 57.3% and 49.6%, respectively. The signature PSTAIRE (single-letter amino acid codes) CDK motif is changed to SKCILRE in PfPK6. The TXY residues that are phosphorylated in MAPKs for their activation are T173PT in PfPK6. Three size classes of PfPK6 transcripts of 6.5, 2.0 and 1.1 kb are up-regulated during the transition of P. falciparum from ring to trophozoite. Western blot analysis suggested the expression of a 35 kDa polypeptide in trophozoites and schizonts. Immunofluorescence studies indicated both nuclear and cytoplasmic localization of PfPK6 in trophozoite, schizont and segmenter stages. In vitro, recombinant PfPK6 phosphorylated itself and also exogenous substrates, histone and the small subunit of the malarial ribonucleotide reductase (R2). The kinase activity of PfPK6 is sensitive to CDK inhibitors such as olomoucine and roscovitine. PfPK6 showed a preference for Mn2+ over Mg2+ ions as a cofactor. The Lys38 → Arg mutant is severely defective in its interaction with ATP and bivalent cations and somewhat defective in catalytic rate for R2 phosphorylation.

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