Cloning and molecular features of cytosolic fructose-1,6-bisphosphatase from pea

2001 ◽  
Vol 28 (2) ◽  
pp. 157
Author(s):  
Rolland Cazalis ◽  
Rolland Cazalis ◽  
Eduardo Pagano ◽  
Eduardo Pagano ◽  
Julio López Gorgé ◽  
...  
Keyword(s):  
Continuous Illumination ◽  
Transcriptional Expression ◽  
Higher Plant ◽  
Molecular Features ◽  
Acid Protein ◽  
Fructose 1,6 Bisphosphatase ◽  
Polymerase Chain ◽  
Expression Mechanism ◽  
Amino Acid Protein ◽  
Cytosolic Fbpase

A cDNA clone encoding for pea (Pisum sativum L.) cytosolic fructose-1,6-bisphosphatase (E.C. 3.1.3.11) has been isolated by reverse transcription-polymerase chain reaction of the total mRNA. The sequence analysis displayed a 341-amino acid protein of about 37300 Da molecular mass, corresponding to the subunit of this homotetrameric enzyme; it showed about 80% homology with the other ten higher plant cytosolic FBPases sequenced so far. The enzyme displayed a strong transcriptional expression in green organs (sessile and petioled leaves, stem, pod and grain), and poor expression in root and senescent basal leaves. It is noteworthy the high FBPase transcriptional expression in pod, which displays up to 4-fold higher content of FBPase-specific mRNA than that of root. The mRNA related to cytosolic FBPase was detected after 24 h continuous illumination of 24-h-dark-grown seedlings; this light-induced transcriptional expression is slower than that of chloroplast FBPase, which appears soon after 2 h light. In both cases the corresponding mRNAs disappeared when the light was turned off. The translational expression was also manifested, both as FBPase protein and activity, after 24 h illumination. This delay in the expression of cytosolic FBPase with respect to that of the plastidic enzyme can be interpreted as an indirect effect induced by a metabolite of the photosynthetic carbon pathway, rather than a direct effect of light on the DNA-expression mechanism. Pea cytosolic FBPase was not activated by dithiothreitol, with or without coupling to thioredoxins f or m. The enzyme showed a half-life of 6 h.

Molecular cloning, characterization, and expression analysis of a novel sema-2a homologue in Polyrhachis vicina (Hymenoptera: Formicidae)

2010 ◽  
Vol 142 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Jing Luo ◽  
Geng-Si Xi ◽  
Shu-Min Lü ◽  
Ke Li ◽  
Qing Li
Keyword(s):  
Untranslated Region ◽  
Axonal Guidance ◽  
Mrna Levels ◽  
Chain Reaction ◽  
Base Pairs ◽  
Reading Frame ◽  
Acid Protein ◽  
Polymerase Chain ◽  
Polyrhachis Vicina ◽  
Amino Acid Protein

AbstractThe semaphorin gene family plays important roles in axonal guidance in vertebrates and invertebrates. Semaphorin 2a, a member of the semaphorin family, belongs to class 2, which is found only in invertebrates. In our study, semaphorin 2a was cloned from the ant Polyrhachis vicina Roger. The full length of P. vicina semaphorin 2a (Pv-sema-2a) is 2763 base pairs (bp) and it contains a 5′-untranslated region (UTR) 92 bp long and a 3′-UTR 521 bp long. The open reading frame of Pv-sema-2a encodes a 716-amino-acid protein with a predicted molecular mass of 81.1 kilodaltons. Real-time quantitative reverse-transcription – polymerase chain reaction indicated that Pv-sema-2a mRNA is differentially expressed during P. vicina development, in the whole bodies as well as the heads of different castes. The high mRNA levels in embryos and pupae suggest that Pv-sema-2a plays an important role in ant development.

scholarly journals Molecular Cloning and Characterization of a cDNA Encoding Sedoheptulose-1,7-bisphosphatase from Mulberry (Morus alba var. multicaulis)

2008 ◽  
Vol 57 (1-6) ◽  
pp. 152-157 ◽  
Author(s):  
X. Ji ◽  
Y. Gai ◽  
J. Ma ◽  
C. Zheng ◽  
Z. Mu
Keyword(s):  
Morus Alba ◽  
Evolutionary Analysis ◽  
Comparison Analysis ◽  
Reading Frame ◽  
Acid Protein ◽  
Fructose 1,6 Bisphosphatase ◽  
Rapid Amplification ◽  
Molecular Evolutionary Analysis ◽  
Amino Acid Protein

Abstract A full-length cDNA encoding sedoheptulose-1,7-bisphosphatase (SBPase; EC 3.1.3.37) was cloned from mulberry (Morus alba var. multicaulis) by rapid amplification of cDNA ends (RACE). The cDNA consisted of 1,527 nucleotides with an open reading frame (ORF) of 1,179 nucleotides encoding a 393 amino acid protein of approximately 42.6 kDa. Sequence comparison analysis showed that mulberry SBPase (MSBPase) had high homology to other plant counterparts. Phylogenetic and molecular evolutionary analysis revealed that MSBPase fell into plant SBPase group. Moreover, SBPase and fructose-1,6-bisphosphatase (FBPase; EC 3.1.3.11) shared 28-32% identical residues, suggesting that the two enzymes originated from the same evolution branch. Molecular modeling indicated that each subunit of MSBPase was composed of α-helices and β-sheets joined by turns and loops, and folded into a structure of hexahedron shape which was very similar to FBPase.

scholarly journals Induction of Cyclo-oxygenase-2 Expression in Naturally Occurring Gastric Ulcers

2002 ◽  
Vol 50 (7) ◽  
pp. 923-933 ◽  
Author(s):  
Stéphane Lajoie ◽  
Jean Sirois ◽  
Monique Doré
Keyword(s):  
Repair Process ◽  
Gastric Ulcers ◽  
Immunoblotting Analysis ◽  
Naturally Occurring ◽  
Acid Protein ◽  
Polymerase Chain ◽  
Cox 2 ◽  
First Time ◽  
Normal Stomach ◽  
Amino Acid Protein

Cyclo-oxygenase-2 (COX-2) is believed to participate in the repair of gastric ulcer. Like humans, pigs frequently develop gastric ulcers and thus represent an attractive animal model in which to study the repair process of naturally occurring gastric ulcers. However, expression of COX in the pig stomach has not been reported. The objectives of this study were to determine whether COX isoenzymes are expressed in porcine gastric ulcers and to characterize the porcine COX-2 cDNA. Normal stomachs ( n=5) and those with gastric ulcers ( n=35) were studied by immunohistochemistry and immunoblotting analysis. Reverse transcription-polymerase chain reaction (RT-PCR) was used to isolate the complete porcine COX-2 cDNA. COX-1 staining was present in normal stomach and in ulcerated areas. No COX-2 was detected in normal stomach, but COX-2 was strongly expressed in the ulcerated area in 28/35 (80%) gastric ulcers ( p<0.01). Immunoblotting analysis confirmed the restricted expression of COX-2 in the ulcerated areas. The porcine COX-2 cDNA was shown to code for a 604 amino acid protein that is 89% identical to human COX-2. These results provide the complete primary structure of porcine COX-2 and demonstrate for the first time that the enzyme is induced in naturally occurring porcine gastric ulcers.

scholarly journals Molecular cloning and sequencing of rabbit presenilin-1 cDNA fragment.

2002 ◽  
Vol 49 (4) ◽  
pp. 1013-1017
Author(s):  
Abdulaziz A Al-Khedhairy ◽  
Misbahul Arfin ◽  
Abdul Aziz Bin Dukhyil
Keyword(s):  
Amino Acid ◽  
Molecular Cloning ◽  
Presenilin 1 ◽  
Cdna Fragment ◽  
Protein Fragment ◽  
Cloning And Sequencing ◽  
Acid Protein ◽  
Polymerase Chain ◽  
Transmembrane Regions ◽  
Amino Acid Protein

Molecular cloning and sequencing of a cDNA encoding rabbit presenilin-1 (Ps1) fragment was performed by reverse transcription polymerase chain reaction (RT-PCR) using primers: 5'-GGA TGA GCA GCT AAT CTA TAC C-3' and 5'-TCC ATT CAG GGA GGT ACT TGA TA-3'. The cDNA fragment revealed 402 nucleotides. The sequence was well conserved and found to be 91, 90, 88, 87 and 78% homologous to that of human, lemur, rat, mouse and chicken, respectively. The cDNA translated into a 130 amino-acid protein fragment. The deduced amino-acid sequence was also well conserved in various species and exhibited 98% similarities with those of rat, lemur and human homologues. However, differences were noticed at residues 145, 168 and 212. This cDNA fragment is quite significant because it is the most conserved portion of Ps1 in various animals and encodes four transmembrane regions (TM2, 3, 4, 5) as defined in human Ps1. Moreover, it includes more than 50% of the sites at which substitutions have been reported in familial Alzheimer's disease (FAD). Therefore, it is suggested that the rabbit can be used as an experimental model for future studies on Ps1 and its physiological functions to work out possible pathways leading to FAD linked neurodegeneration.

scholarly journals Characterization of the Gallate Dioxygenase Gene: Three Distinct Ring Cleavage Dioxygenases Are Involved in Syringate Degradation by Sphingomonas paucimobilis SYK-6

2005 ◽  
Vol 187 (15) ◽  
pp. 5067-5074 ◽  
Author(s):  
Daisuke Kasai ◽  
Eiji Masai ◽  
Keisuke Miyauchi ◽  
Yoshihiro Katayama ◽  
Masao Fukuda
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequences ◽  
Terminal Region ◽  
Ring Cleavage ◽  
Sphingomonas Paucimobilis ◽  
Acid Protein ◽  
Demethylation Pathway ◽  
Dioxygenase Gene ◽  
Amino Acid Protein

ABSTRACT Sphingomonas paucimobilis SYK-6 converts vanillate and syringate to protocatechuate (PCA) and 3-O-methylgallate (3MGA) in reactions with the tetrahydrofolate-dependent O-demethylases LigM and DesA, respectively. PCA is further degraded via the PCA 4,5-cleavage pathway, whereas 3MGA is metabolized via three distinct pathways in which PCA 4,5-dioxygenase (LigAB), 3MGA 3,4-dioxygenase (DesZ), and 3MGA O-demethylase (LigM) are involved. In the 3MGA O-demethylation pathway, LigM converts 3MGA to gallate, and the resulting gallate appears to be degraded by a dioxygenase other than LigAB or DesZ. Here, we isolated the gallate dioxygenase gene, desB, which encodes a 418-amino-acid protein with a molecular mass of 46,843 Da. The amino acid sequences of the N-terminal region (residues 1 to 285) and the C-terminal region (residues 286 to 418) of DesB exhibited ca. 40% and 27% identity with the sequences of the PCA 4,5-dioxygenase β and α subunits, respectively. DesB produced in Escherichia coli was purified and was estimated to be a homodimer (86 kDa). DesB specifically attacked gallate to generate 4-oxalomesaconate as the reaction product. The Km for gallate and the V max were determined to be 66.9 ± 9.3 μM and 42.7 ± 2.4 U/mg, respectively. On the basis of the analysis of various SYK-6 mutants lacking the genes involved in syringate degradation, we concluded that (i) all of the three-ring cleavage dioxygenases are involved in syringate catabolism, (ii) the pathway involving LigM and DesB plays an especially important role in the growth of SYK-6 on syringate, and (iii) DesB and LigAB are involved in gallate degradation.

scholarly journals LASS3 (longevity assurance homologue 3) is a mainly testis-specific (dihydro)ceramide synthase with relatively broad substrate specificity

2006 ◽  
Vol 398 (3) ◽  
pp. 531-538 ◽  
Author(s):  
Yukiko Mizutani ◽  
Akio Kihara ◽  
Yasuyuki Igarashi
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Family Members ◽  
Fatty Acyl ◽  
Ceramide Synthase ◽  
Broad Substrate Specificity ◽  
Acid Protein ◽  
Amino Acid Protein

The LASS (longevity assurance homologue) family members are highly conserved from yeasts to mammals. Five mouse and human LASS family members, namely LASS1, LASS2, LASS4, LASS5 and LASS6, have been identified and characterized. In the present study we cloned two transcriptional variants of hitherto-uncharacterized mouse LASS3 cDNA, which encode a 384-amino-acid protein (LASS3) and a 419-amino-acid protein (LASS3-long). In vivo, [3H]dihydrosphingosine labelling and electrospray-ionization MS revealed that overproduction of either LASS3 isoform results in increases in several ceramide species, with some preference toward those having middle- to long-chain-fatty acyl-CoAs. A similar substrate preference was observed in an in vitro (dihydro)ceramide synthase assay. These results indicate that LASS3 possesses (dihydro)ceramide synthesis activity with relatively broad substrate specificity. We also found that, except for a weak display in skin, LASS3 mRNA expression is limited almost solely to testis, implying that LASS3 plays an important role in this gland.

Isolation, DNA sequence, and regulation of a Saccharomyces cerevisiae gene that encodes DNA strand transfer protein alpha

1991 ◽  
Vol 11 (5) ◽  
pp. 2576-2582
Author(s):  
A B Clark ◽  
C C Dykstra ◽  
A Sugino
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Specific Activity ◽  
Intragenic Recombination ◽  
Strand Transfer ◽  
Homologous Pairing ◽  
Spore Viability ◽  
Transfer Protein ◽  
Acid Protein ◽  
Dna Strand ◽  
Amino Acid Protein

DNA strand transfer protein alpha (STP alpha) from meiotic Saccharomyces cerevisiae cells promotes homologous pairing of DNA without any nucleotide cofactor in the presence of yeast single-stranded DNA binding protein. This gene (DNA strand transferase 1, DST1) encodes a 309-amino-acid protein with a predicted molecular mass of 34,800 Da. The STP alpha protein level is constant in both mitotic and meiotic cells, but during meiosis the polypeptide is activated by an unknown mechanism, resulting in a large increase in its specific activity. A dst1::URA3/dst1::URA3 mutant grows normally in mitotic media; however, meiotic cells exhibit a greatly reduced induction of both DNA strand transfer activity and intragenic recombination between his1 heteroalleles. Spore viability is normal. These results suggest that DST1 is required for much of the observed induction of homologous recombination in S. cerevisiae during meiosis but not for normal sporulation.

glsA, a Volvox gene required for asymmetric division and germ cell specification, encodes a chaperone-like protein

Development ◽  
1999 ◽  
Vol 126 (4) ◽  
pp. 649-658 ◽  
Author(s):  
S.M. Miller ◽  
D.L. Kirk
Keyword(s):  
Mitotic Spindle ◽  
Site Directed Mutagenesis ◽  
Cell Specification ◽  
Division Plane ◽  
Binding Domains ◽  
Acid Protein ◽  
Germ Cell Specification ◽  
Asymmetric Divisions ◽  
Dividing Cells ◽  
Amino Acid Protein

The gls genes of Volvox are required for the asymmetric divisions that set apart cells of the germ and somatic lineages during embryogenesis. Here we used transposon tagging to clone glsA, and then showed that it is expressed maximally in asymmetrically dividing embryos, and that it encodes a 748-amino acid protein with two potential protein-binding domains. Site-directed mutagenesis of one of these, the J domain (by which Hsp40-class chaperones bind to and activate specific Hsp70 partners) abolishes the capacity of glsA to rescue mutants. Based on this and other considerations, including the fact that the GlsA protein is associated with the mitotic spindle, we discuss how it might function, in conjunction with an Hsp70-type partner, to shift the division plane in asymmetrically dividing cells.

scholarly journals JAZ repressors of metabolic defense promote growth and reproductive fitness inArabidopsis

2018 ◽  
Vol 115 (45) ◽  
pp. E10768-E10777 ◽  
Author(s):  
Qiang Guo ◽  
Yuki Yoshida ◽  
Ian T. Major ◽  
Kun Wang ◽  
Koichi Sugimoto ◽  
...  
Keyword(s):  
Gene Family ◽  
Fungal Pathogens ◽  
Defense Responses ◽  
Protein Profiling ◽  
Metabolic Effects ◽  
Repressor Proteins ◽  
Acid Protein ◽  
Jaz Proteins ◽  
Physiological Tasks ◽  
Amino Acid Protein

Plant immune responses mediated by the hormone jasmonoyl-l-isoleucine (JA-Ile) are metabolically costly and often linked to reduced growth. Although it is known that JA-Ile activates defense responses by triggering the degradation of JASMONATE ZIM DOMAIN (JAZ) transcriptional repressor proteins, expansion of theJAZgene family in vascular plants has hampered efforts to understand how this hormone impacts growth and other physiological tasks over the course of ontogeny. Here, we combined mutations within the 13-memberArabidopsis JAZgene family to investigate the effects of chronic JAZ deficiency on growth, defense, and reproductive output. A higher-order mutant (jazdecuple,jazD) defective in 10JAZgenes (JAZ1–7,-9,-10, and-13) exhibited robust resistance to insect herbivores and fungal pathogens, which was accompanied by slow vegetative growth and poor reproductive performance. Metabolic phenotypes ofjazDdiscerned from global transcript and protein profiling were indicative of elevated carbon partitioning to amino acid-, protein-, and endoplasmic reticulum body-based defenses controlled by the JA-Ile and ethylene branches of immunity. Resource allocation to a strong defense sink injazDleaves was associated with increased respiration and hallmarks of carbon starvation but no overt changes in photosynthetic rate. Depletion of the remaining JAZ repressors injazDfurther exaggerated growth stunting, nearly abolished seed production and, under extreme conditions, caused spreading necrotic lesions and tissue death. Our results demonstrate that JAZ proteins promote growth and reproductive success at least in part by preventing catastrophic metabolic effects of an unrestrained immune response.

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