Expression of a UDPglucose pyrophosphorylase cDNA during fruit ripening of banana (Musa acuminata)

2000 ◽  
Vol 27 (12) ◽  
pp. 1151 ◽  
Author(s):  
Eng-Chong Pua ◽  
Stevenson Szu-Wei Lim ◽  
Pei Liu ◽  
Jian-Zhong Liu
Keyword(s):  
Sequence Similarity ◽  
Soluble Sugars ◽  
Transcript Abundance ◽  
Reproductive Organs ◽  
Banana Fruit ◽  
Amino Acid Residues ◽  
Promoting Effect ◽  
Udpglucose Pyrophosphorylase ◽  
Exogenous Ethylene ◽  
High Degree

We report the isolation of a banana cDNA, designated MWUGPA, encoding uridine diphosphoryl (UDP)-glucose pyrophosphorylase (UGPase, EC.2.7.7.9) that catalyses the reversible conversion between glucose 1-phosphate and UDPglucose in plants and animals. Furthermore, UGPase expression in fruit during ripening and in response to exogenous ethylene and sugars was also investigated. MWUGPA encodes a polypeptide of 467 amino acid residues and shares a high degree of sequence similarity (85–90%) with other plant UGPase homologs. In northern blot analysis, a 1.7-kb UGPase transcript was detected in both the vegetative and reproductive organs, but the former was considerably less abundant than the latter. In fruit, the level of accumulated transcripts was higher in pulp than peel at all ripening stages. Transcript abundance in both fruit tissues was relatively constant during ripen-ing, but pulp transcripts surged in the ‘more green than yellow’ category fruit when ethylene also increased. Further analysis revealed that UGPase expression in fruit was ethylene-inducible, but the response was tissue-specific, as evidenced by the promoting effect of exogenous ethylene on accumulation of UGPase transcripts in pulp but not peel. Exogenous application of sucrose and fructose also increased UGPase transcript abundance in leaf and fruit tissues, especially pulp, whereas exogenous glucose had little or no effect. The results of this study indicate that ethy-lene and soluble sugars may play a regulatory role in UGPase expression during ripening in banana fruit.

scholarly journals The ς70 Transcription Factor TyrR Has Zinc-Stimulated Phosphatase Activity That Is Inhibited by ATP and Tyrosine

2000 ◽  
Vol 182 (4) ◽  
pp. 1053-1061 ◽  
Author(s):  
Shimin Zhao ◽  
Qin Zhu ◽  
Ronald L. Somerville
Keyword(s):  
Amino Acid ◽  
Phosphatase Activity ◽  
Sequence Similarity ◽  
Zinc Ion ◽  
Amino Acid Residues ◽  
Protein Residues ◽  
E Coli ◽  
C Terminus ◽  
Atp Analogues ◽  
High Degree

ABSTRACT The TyrR protein of Escherichia coli (513 amino acid residues) is the chief transcriptional regulator of a group of genes that are essential for aromatic amino acid biosynthesis and transport. The TyrR protein can function either as a repressor or as an activator. The central region of the TyrR protein (residues 207 to 425) is similar to corresponding polypeptide segments of the NtrC protein superfamily. Like the NtrC protein, TyrR has intrinsic ATPase activity. Here, we report that TyrR possesses phosphatase activity. This activity is subject to inhibition by l-tyrosine and its analogues and by ATP and ATP analogues. Zinc ion (2 mM) stimulated the phosphatase activity of the TyrR protein by a factor of 57. The phosphatase-active site of TyrR was localized to a 31-kDa domain (residues 191 to 467) of the protein. However, mutational alteration of distant amino acid residues at both the N terminus and the C terminus of TyrR altered the phosphatase activity. Haemophilus influenzae TyrR (318 amino acid residues), a protein with a high degree of sequence similarity to the C terminus of the E. coli TyrR protein, exhibited a phosphatase activity similar to that of E. coliTyrR.

scholarly journals Molecular cloning and sequence analysis of the cDNA for ancrod, a thrombin-like enzyme from the venom of Calloselasma rhodostoma

1993 ◽  
Vol 294 (2) ◽  
pp. 387-390 ◽  
Author(s):  
L C Au ◽  
S B Lin ◽  
J S Chou ◽  
G W Teh ◽  
K J Chang ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Serine Proteases ◽  
Sequence Similarity ◽  
Active Form ◽  
Amino Acid Sequences ◽  
Amino Acid Residues ◽  
Binding Reaction ◽  
Venom Glands ◽  
High Degree

The 1.54 kb cDNA for ancrod, a thrombin-like enzyme, was cloned from a lambda ZAP cDNA library derived from the venom glands of Calloselasma (Agkistrodon) rhodostoma. The cDNA sequence reveals that ancrod is synthesized as a pre-zymogen of 258 amino acids, including a putative secretory peptide of 18 amino acids and a proposed zymogen peptide of 6 amino-acid residues. The amino-acid sequence of the predicted active form of the enzyme exhibits a high degree of sequence similarity to those of mammalian serine proteases (trypsin and pancreatic kallikrein) and other thrombin-like enzymes (batroxobin and flavoxobin). Key amino-acid residues (His43, Asp88, Ser182 and Asp176) that are thought to be involved in the substrate cleavage and in the substrate-binding reaction are conserved. Ancrod contains 13 cysteine residues. Based on alignment with the amino-acid sequences of trypsin and batroxobin, six disulphide bridges can be predicted to be present in the ancrod protein. The existence of a free cysteine, which changes the common sequence surrounding the Ser182 active site from Gly-Asp-Ser-Gly-Gly-Pro to Cys-Asp-Ser-Gly-Gly-Pro, is unusual for a serine protease.

scholarly journals Structural and functional analysis of the Na+/H+ exchanger

2007 ◽  
Vol 401 (3) ◽  
pp. 623-633 ◽  
Author(s):  
Emily R. Slepkov ◽  
Jan K. Rainey ◽  
Brian D. Sykes ◽  
Larry Fliegel
Keyword(s):  
Intracellular Ph ◽  
Sequence Similarity ◽  
Structural Data ◽  
Structural Similarity ◽  
Integral Membrane Protein ◽  
Volume Control ◽  
Amino Acid Residues ◽  
Physiological Processes ◽  
High Resolution Structure ◽  
Extracellular Sodium

The mammalian NHE (Na+/H+ exchanger) is a ubiquitously expressed integral membrane protein that regulates intracellular pH by removing a proton in exchange for an extracellular sodium ion. Of the nine known isoforms of the mammalian NHEs, the first isoform discovered (NHE1) is the most thoroughly characterized. NHE1 is involved in numerous physiological processes in mammals, including regulation of intracellular pH, cell-volume control, cytoskeletal organization, heart disease and cancer. NHE comprises two domains: an N-terminal membrane domain that functions to transport ions, and a C-terminal cytoplasmic regulatory domain that regulates the activity and mediates cytoskeletal interactions. Although the exact mechanism of transport by NHE1 remains elusive, recent studies have identified amino acid residues that are important for NHE function. In addition, progress has been made regarding the elucidation of the structure of NHEs. Specifically, the structure of a single TM (transmembrane) segment from NHE1 has been solved, and the high-resolution structure of the bacterial Na+/H+ antiporter NhaA has recently been elucidated. In this review we discuss what is known about both functional and structural aspects of NHE1. We relate the known structural data for NHE1 to the NhaA structure, where TM IV of NHE1 shows surprising structural similarity with TM IV of NhaA, despite little primary sequence similarity. Further experiments that will be required to fully understand the mechanism of transport and regulation of the NHE1 protein are discussed.

scholarly journals Cloning, post-translational modifications, heterologous expression and ligand-binding of boar salivary lipocalin

2000 ◽  
Vol 350 (2) ◽  
pp. 369-379 ◽  
Author(s):  
Dietrich LOEBEL ◽  
Andrea SCALONI ◽  
Sara PAOLINI ◽  
Carlo FINI ◽  
Lino FERRARA ◽  
...  
Keyword(s):  
Amino Acid ◽  
Sequence Similarity ◽  
Three Dimensional ◽  
Protein Isoforms ◽  
Cysteine Residues ◽  
Protein Chemistry ◽  
Single Individual ◽  
Amino Acid Residues ◽  
Three Dimensional Model ◽  
Submaxillary Glands

Boar submaxillary glands produce the sex-specific salivary lipocalin (SAL), which binds steroidal sex pheromones as endogenous ligands. The cDNA encoding SAL was cloned and sequenced. From a single individual, two protein isoforms, differing in three amino acid residues, were purified and structurally characterized by a combined Edman degradation/MS approach. These experiments ascertained that the mature polypeptide is composed of 168 amino acid residues, that one of the three putative glycosylation sites is post-translationally modified and the structure of the bound glycosidic moieties. Two of the cysteine residues are paired together in a disulphide bridge, whereas the remaining two occur as free thiols. SAL bears sequence similarity to other lipocalins; on this basis, a three-dimensional model of the protein has been built. A SAL isoform was expressed in Escherichiacoli in good yields. Protein chemistry and CD experiments verified that the recombinant product shows the same redox state at the cysteine residues and that the same conformation is observed as in the natural protein, thus suggesting similar folding. Binding experiments on natural and recombinant SAL were performed with the fluorescent probe 1-aminoanthracene, which was efficiently displaced by the steroidal sex pheromone, as well as by several odorants.

scholarly journals The TFIID Components Human TAFII140 andDrosophila BIP2 (TAFII155) Are Novel Metazoan Homologues of Yeast TAFII47 Containing a Histone Fold and a PHD Finger

2001 ◽  
Vol 21 (15) ◽  
pp. 5109-5121 ◽  
Author(s):  
Yann-Gaël Gangloff ◽  
Jean-Christophe Pointud ◽  
Sylvie Thuault ◽  
Lucie Carré ◽  
Christophe Romier ◽  
...  
Keyword(s):  
Rna Polymerase Ii ◽  
Sequence Similarity ◽  
Protein A ◽  
Amino Acid Sequences ◽  
Molecular Organization ◽  
Phd Finger ◽  
Histone Fold ◽  
Histone Fold Domain ◽  
High Degree

ABSTRACT The RNA polymerase II transcription factor TFIID comprises the TATA binding protein (TBP) and a set of TBP-associated factors (TAFIIs). TFIID has been extensively characterized for yeast, Drosophila, and humans, demonstrating a high degree of conservation of both the amino acid sequences of the constituent TAFIIs and overall molecular organization. In recent years, it has been assumed that all the metazoan TAFIIs have been identified, yet no metazoan homologues of yeast TAFII47 (yTAFII47) and yTAFII65 are known. Both of these yTAFIIs contain a histone fold domain (HFD) which selectively heterodimerizes with that of yTAFII25. We have cloned a novel mouse protein, TAFII140, containing an HFD and a plant homeodomain (PHD) finger, which we demonstrated by immunoprecipitation to be a mammalian TFIID component. TAFII140 shows extensive sequence similarity toDrosophila BIP2 (dBIP2) (dTAFII155), which we also show to be a component of DrosophilaTFIID. These proteins are metazoan homologues of yTAFII47 as their HFDs selectively heterodimerize with dTAFII24 and human TAFII30, metazoan homologues of yTAFII25. We further show that yTAFII65 shares two domains with theDrosophila Prodos protein, a recently described potential dTAFII. These conserved domains are critical for yTAFII65 function in vivo. Our results therefore identify metazoan homologues of yTAFII47 and yTAFII65.

scholarly journals Action of ethylene on postharvest of summer squash ‘Menina Brasileira’

Revista CERES ◽  
2017 ◽  
Vol 64 (4) ◽  
pp. 360-367
Author(s):  
Fernanda Ferreira de Araújo ◽  
Lucas Cavalcante da Costa ◽  
Tania Pires da Silva ◽  
Mário Puiatti ◽  
Fernando Luiz Finger
Keyword(s):  
Weight Loss ◽  
Chlorophyll Content ◽  
Electrolyte Leakage ◽  
Soluble Sugars ◽  
Control Treatment ◽  
Ethylene Concentration ◽  
Malondialdehyde Content ◽  
Summer Squash ◽  
No Weight Loss ◽  
Exogenous Ethylene

ABSTRACT The purpose of this study was to evaluate the sensitivity and the physiological responses of summer squash ‘Menina Brasileira’ to ethylene. Immature fruits were harvested and placed in 20 L sealed buckets, in which ethylene was applied at concentrations of 0.1, 1.0, 10, 100, and 1000 μL L-1 for 24 h. Fruits were placed in buckets with no ethylene as a control treatment. Thereafter, the fruits were taken out of the buckets and maintained on bench, wherein on days 0, 2, 4, 6, and 8, they were evaluated regarding the accumulated fresh weight loss, soluble sugars, reducing and non-reducing sugars, starch, total chlorophyll, content of malondialdehyde, and electrolyte leakage. Fruits of summer squash ‘Menina Brasileira’ showed sensitivity to exogenous ethylene with no weight loss stimulation. Additionally, the fruits exhibited small changes in nutritional quality attributes and changes in the external fruit appearance, including decreased chlorophyll content as well as damage to cell membrane characterized by increase in malondialdehyde content and electrolyte leakage. These changes were stimulated by increasing exogenous ethylene concentration.

scholarly journals Isolation and amino-acid sequence of two inhibitors of serine proteinases, members of the squash inhibitor family, from Echinocystis lobata seeds.

1996 ◽  
Vol 43 (3) ◽  
pp. 507-513 ◽  
Author(s):  
D Stachowiak ◽  
A Polanowski ◽  
G Bieniarz ◽  
T Wilusz
Keyword(s):  
Amino Acid ◽  
Proteinase Inhibitors ◽  
Sequence Similarity ◽  
Serine Proteinase ◽  
Exchange Chromatography ◽  
Association Constants ◽  
Cathepsin G ◽  
Amino Acid Residues ◽  
Mature Seeds ◽  
Echinocystis Lobata

Two serine proteinase inhibitors (ELTI I and ELTI II) have been isolated from mature seeds of Echinocystis lobata by ammonium sulfate fractionation, methanol precipitation, ion exchange chromatography, affinity chromatography on immobilized anhydrotrypsin and HPLC. ELTI I and ELTI II consist of 33 and 29 amino-acid residues, respectively. The primary structures of these inhibitors are as follows: ELTI I KEEQRVCPRILMRCKRDSDCLAQCTCQQSGFCG ELTI II RVCPRILMRCKRDSDCLAQCTCQQSGFCG The inhibitors show sequence similarity with the squash inhibitor family. ELTI I differs from ELTI II only by the presence of the NH2-terminal tetrapeptide Lys-Glu-Glu-Gln. The association constants (Ka) of ELTI I and ELTI II with bovine-trypsin were determined to be 6.6 x 10(10) M-1, and 3.1 x 10(11) M-1, whereas the association constants of these inhibitors with cathepsin G were 1.2 x 10(7) M-1, and 1.1 x 10(7) M-1, respectively.

scholarly journals Nitrogen Fixation Genes in an EndosymbioticBurkholderia Strain

2001 ◽  
Vol 67 (2) ◽  
pp. 725-732 ◽  
Author(s):  
Daniela Minerdi ◽  
Renato Fani ◽  
Romina Gallo ◽  
Alessandra Boarino ◽  
Paola Bonfante
Keyword(s):  
Genomic Library ◽  
Sequence Similarity ◽  
Bacterial Genome ◽  
Fungal Spores ◽  
Mycorrhizal Fungus ◽  
Prokaryotic Genome ◽  
Arbuscular Mycorrhizal ◽  
Open Reading Frames ◽  
Genome Screening ◽  
High Degree

ABSTRACT In this paper we report the identification and characterization of a DNA region containing putative nif genes and belonging to a Burkholderia endosymbiont of the arbuscular mycorrhizal fungus Gigaspora margarita. A genomic library of total DNA extracted from the fungal spores was also representative of the bacterial genome and was used to investigate the prokaryotic genome. Screening of the library with Azospirillum brasilense nifHDKgenes as the prokaryotic probes led to the identification of a 6,413-bp region. Analysis revealed three open reading frames encoding putative proteins with a very high degree of sequence similarity with the two subunits (NifD and NifK) of the component I and with component II (NifH) of nitrogenase from different diazotrophs. The three genes were arranged in an operon similar to that shown by most archaeal and bacterial diazotrophs. PCR experiments with primers designed on theBurkholderia nifHDK genes and Southern blot analysis demonstrate that they actually belong to the genome of the G. margarita endosymbiont. They offer, therefore, the first sequence for the nif operon described for Burkholderia. Reverse transcriptase PCR experiments with primers designed on theBurkholderia nifH and nifD genes and performed on total RNA extracted from spores demonstrate that the gene expression was limited to the germination phase. A phylogenetic analysis performed on the available nifK sequences placed the endosymbioticBurkholderia close to A. brasilense.

Remarkable intron and exon sequence conservation in human and mouse homeobox Hox 1.3 genes

1989 ◽  
Vol 9 (5) ◽  
pp. 2273-2278
Author(s):  
E Tournier-Lasserve ◽  
W F Odenwald ◽  
J Garbern ◽  
J Trojanowski ◽  
R A Lazzarini
Keyword(s):  
Sequence Similarity ◽  
Gene Clusters ◽  
Binding Studies ◽  
Base Pairs ◽  
Cis Element ◽  
Multiple Transcripts ◽  
Exon Sequence ◽  
Dna Binding Studies ◽  
High Degree ◽  
Sequence Similarities

A high degree of conservation exists between the Hox 1.3 homeobox genes of mice and humans. The two genes occupy the same relative positions in their respective Hox 1 gene clusters, they show extensive sequence similarities in their coding and noncoding portions, and both are transcribed into multiple transcripts of similar sizes. The predicted human Hox 1.3 protein differs from its murine counterpart in only 7 of 270 amino acids. The sequence similarity in the 250 base pairs upstream of the initiation codon is 98%, the similarity between the two introns, both 960 base pairs long, is 72%, and the similarity in the 3' noncoding region from termination codon to polyadenylation signal is 90%. Both mouse and human Hox 1.3 introns contain a sequence with homology to a mating-type-controlled cis element of the yeast Ty1 transposon. DNA-binding studies with a recombinant mouse Hox 1.3 protein identified two binding sites in the intron, both of which were within the region of shared homology with this Ty1 cis element.

The FHA domain mediates phosphoprotein interactions

2000 ◽  
Vol 113 (23) ◽  
pp. 4143-4149 ◽  
Author(s):  
J. Li ◽  
G.I. Lee ◽  
S.R. Van Doren ◽  
J.C. Walker
Keyword(s):  
Sequence Similarity ◽  
Sequence Motif ◽  
Amino Acid Residues ◽  
Fha Domain ◽  
Tertiary Structures ◽  
Cellular Processes ◽  
Forkhead Transcription Factors ◽  
Binding Domains ◽  
Cycle Arrest ◽  
Protein Kinase Signaling

The forkhead-associated (FHA) domain is a phosphopeptide-binding domain first identified in a group of forkhead transcription factors but is present in a wide variety of proteins from both prokaryotes and eukaryotes. In yeast and human, many proteins containing an FHA domain are found in the nucleus and involved in DNA repair, cell cycle arrest, or pre-mRNA processing. In plants, the FHA domain is part of a protein that is localized to the plasma membrane and participates in the regulation of receptor-like protein kinase signaling pathways. Recent studies show that a functional FHA domain consists of 120–140 amino acid residues, which is significantly larger than the sequence motif first described. Although FHA domains do not exhibit extensive sequence similarity, they share similar secondary and tertiary structures, featuring a sandwich of two anti-parallel (beta)-sheets. One intriguing finding is that FHA domains may bind phosphothreonine, phosphoserine and sometimes phosphotyrosine, distinguishing them from other well-studied phosphoprotein-binding domains. The diversity of proteins containing FHA domains and potential differences in binding specificities suggest the FHA domain is involved in coordinating diverse cellular processes.

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