scholarly journals S-Like Ribonuclease T2 Genes Are Induced during Mobilisation of Nutrients in Cotyledons from Common Bean

Agronomy ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 490
Author(s):  
Mercedes Diaz-Baena ◽  
Elena Delgado-García ◽  
Manuel Pineda ◽  
Gregorio Galvez-Valdivieso ◽  
Pedro Piedras
Keyword(s):  
Common Bean ◽  
Physiological Function ◽  
Specific Activity ◽  
Seed Viability ◽  
Seedling Development ◽  
Ribonuclease Activity ◽  
Rna Turnover ◽  
Regulatory Signal ◽  
Molecular Masses ◽  
Nutrient Mobilisation

Germination and seedling development are crucial phases in a plant’s life cycle with economical and agronomical implications. The RNA quality in seeds is linked to seed viability, being an important agronomic trait since this leads to a loss in germination efficiency. In addition, RNA can be an important phosphorous reservoir in seeds, affecting the efficiency of the mobilisation of nutrients towards the seedlings. However, knowledge about the physiological function of ribonucleases during germination and seedling development is scarce. We analysed the ribonuclease activities of cotyledons during these processes and the expression of S-like ribonucleases T2. Ribonuclease activity was detected in cotyledons at 1 day after imbibition and the specific activity increased during germination and seedling development, reaching a maximal value at 10 days after imbibition. At this stage, the levels of proteins and RNA in cotyledons were very low. Using in-gel assays, three ribonucleases were detected with apparent molecular masses of 16, 17 and 19 kDa along cotyledon ontogeny. The S-like ribonucleases T2 family consists of four genes in common bean (PvRNS1 to PvRNS4). The expression of PvRNS1, PvRNS2 and PvRNS4 increased in the phase of nutrient mobilisation in cotyledons. The expression of PvRNS1 increased 1000 fold in cotyledons, from 1 to 6 days after imbibition. The suppression of the induction of ribonuclease activity and gene expression in decapitated seedlings suggests that the regulatory signal comes from the developing axes. These results clearly state that S-like ribonucleases T2 are involved in RNA turnover in cotyledons during seedling development.

scholarly journals Seed germination at different stratification temperatures and development of Phytelephas macrocarpa Ruiz & Pavón seedlings

2017 ◽  
Vol 39 (1) ◽  
pp. 20-26 ◽  
Author(s):  
Sidney Alberto do Nascimento Ferreira ◽  
Daniel Felipe de Oliveira Gentil
Keyword(s):  
Seed Germination ◽  
Seed Dormancy ◽  
Seed Viability ◽  
Seedling Development ◽  
Development Stages ◽  
Plastic Bags ◽  
Alternating Temperature

Abstract: Phytelephas macrocarpa (ivory palm) is an Amazonian palm vulnerable to exploitation pressure, as its seeds are widely used in regional handicrafts. The aims of this study were to evaluate the effectiveness of different stratification temperatures in overcoming seed dormancy and to analyze the seedling development stages of this species. In germination under stratification, the seeds were placed in plastic bags containing moistened vermiculite, and maintained at the constant temperatures of 25, 30, 35 and 40 °C, and an alternating temperature from 26 to 40 °C. In the study about the development of seedlings, seeds were sown in vermiculite under plastic cover (growing house), and the evolution of the seedling was evaluated, from the formation of germinative button to the complete blade expansion of the first eophyll. Stratification at alternating temperatures (26 to 40 °C) helped overcoming seed dormancy. Stratification at 25 °C kept the seed viability for nine months. Germination, characterized by the formation of the germinative button took an average of 114 ± 24 days, and the seedling development until the first extended eophyll lasted 244 ± 57 days.

RIBONUCLEASE ACTIVITY IN RUSTED WHEAT LEAVES

1961 ◽  
Vol 39 (4) ◽  
pp. 775-784 ◽  
Author(s):  
R. Rohringer ◽  
D. J. Samborski ◽  
C. O. Person
Keyword(s):  
Leaf Rust ◽  
Direct Evidence ◽  
Specific Activity ◽  
Host Tissue ◽  
Ribonuclease Activity ◽  
Rnase Activity ◽  
Rnase Inhibitor ◽  
Germination Medium ◽  
Wheat Leaves ◽  
Host Tissues

Extracts from primary leaves of Lee wheat were prepared at various days following inoculation with races of leaf rust and tested for ribonuclease (RNase) activity. As early as 24 hours after inoculation there was a marked increase in the specific activity of the enzyme in extracts of rusted host tissues. A further increase in activity was observed during later stages of infection, with the susceptible and resistant reacting tissue differing only in the degree of their response. Extracts from noninoculated control leaves exhibited a constant RNase activity throughout the period of observation. The germination medium and extracts from germinating uredospores contained comparatively little RNase activity. No direct evidence was obtained either for the possible release of the enzyme from particulate cellular fractions of the host tissue as a result of infection or for the removal of an RNase inhibitor in the host tissue responding to infection.

scholarly journals Characterization of an Exceedingly Active NADH Oxidase from the Anaerobic Hyperthermophilic Bacterium Thermotoga maritima

2007 ◽  
Vol 189 (8) ◽  
pp. 3312-3317 ◽  
Author(s):  
Xianqin Yang ◽  
Kesen Ma
Keyword(s):  
Nadh Oxidase ◽  
Specific Activity ◽  
Ph Value ◽  
Thermotoga Maritima ◽  
Cell Extract ◽  
Hyperthermophilic Bacterium ◽  
Its Gene ◽  
Molecular Masses ◽  
New Type

ABSTRACT An NADH oxidase from the anaerobic hyperthermophilic bacterium Thermotoga maritima was purified. The enzyme was very active in catalyzing the reduction of oxygen to hydrogen peroxide with an optimal pH value of 7 at 80°C. The Vmax was 230 ± 14 μmol/min/mg (k cat/Km = 548,000 min−1 mM−1), and the Km values for NADH and oxygen were 42 ± 3 and 43 ± 4 μM, respectively. The NADH oxidase was a heterodimeric flavoprotein with two subunits with molecular masses of 54 kDa and 46 kDa. Its gene sequences were identified, and the enzyme might represent a new type of NADH oxidase in anaerobes. An NADH-dependent peroxidase with a specific activity of 0.1 U/mg was also present in the cell extract of T. maritima.

scholarly journals Collateral fitness effects of mutations

2020 ◽  
Vol 117 (21) ◽  
pp. 11597-11607 ◽  
Author(s):  
Jacob D. Mehlhoff ◽  
Frank W. Stearns ◽  
Dahlia Rohm ◽  
Buheng Wang ◽  
Erh-Yeh Tsou ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Protein Evolution ◽  
Physiological Function ◽  
Specific Activity ◽  
Antibiotic Resistance Gene ◽  
Cell Phenotype ◽  
Missense Mutations ◽  
Protein Activity ◽  
Fitness Effects ◽  
As Effects

The distribution of fitness effects of mutation plays a central role in constraining protein evolution. The underlying mechanisms by which mutations lead to fitness effects are typically attributed to changes in protein specific activity or abundance. Here, we reveal the importance of a mutation’s collateral fitness effects, which we define as effects that do not derive from changes in the protein’s ability to perform its physiological function. We comprehensively measured the collateral fitness effects of missense mutations in theEscherichia coli TEM-1β-lactamase antibiotic resistance gene using growth competition experiments in the absence of antibiotic. At least 42% of missense mutations inTEM-1were deleterious, indicating that for some proteins collateral fitness effects occur as frequently as effects on protein activity and abundance. Deleterious mutations caused improper posttranslational processing, incorrect disulfide-bond formation, protein aggregation, changes in gene expression, and pleiotropic effects on cell phenotype. Deleterious collateral fitness effects occurred more frequently inTEM-1than deleterious effects on antibiotic resistance in environments with low concentrations of the antibiotic. The surprising prevalence of deleterious collateral fitness effects suggests they may play a role in constraining protein evolution, particularly for highly expressed proteins, for proteins under intermittent selection for their physiological function, and for proteins whose contribution to fitness is buffered against deleterious effects on protein activity and protein abundance.

scholarly journals Purification and characterization of rat liver glycosylasparaginase

1989 ◽  
Vol 260 (1) ◽  
pp. 101-108 ◽  
Author(s):  
O K Tollersrud ◽  
N N Aronson
Keyword(s):  
Rat Liver ◽  
Gel Electrophoresis ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Polyacrylamide Gel ◽  
Oligosaccharide Chain ◽  
Molecular Masses ◽  
High Mannose

1. Rat liver glycosylasparaginase [N4-(beta-N-acetylglucosaminyl)-L-asparaginase, EC 3.5.1.26] was purified to homogeneity by using salt fractionation, CM-cellulose and DEAE-cellulose chromatography, gel filtration on Ultrogel AcA-54, concanavalin A-Sepharose affinity chromatography, heat treatment at 70 degrees C and preparative SDS/polyacrylamide-gel electrophoresis. The purified enzyme had a specific activity of 3.8 mumol of N-acetylglucosamine/min per mg with N4-(beta-N-acetylglucosaminyl)-L-asparagine as substrate. 2. The native enzyme had a molecular mass of 49 kDa and was composed of two non-identical subunits joined by strong non-covalent forces and having molecular masses of 24 and 20 kDa as determined by SDS/polyacrylamide-gel electrophoresis. 3. The 20 kDa subunit contained one high-mannose-type oligosaccharide chain, and the 24 kDa subunit had one high-mannose-type and one complex-type oligosaccharide chain. 4. N-Terminal sequence analysis of each subunit revealed a frayed N-terminus of the 24 kDa subunit and an apparent N-glycosylation of Asn-15 in the same subunit. 5. The enzyme exhibited a broad pH maximum above 7. Two major isoelectric forms were found at pH 6.4 and 6.6. 6. Glycosylasparaginase was stable at 75 degrees C and in 5% (w/v) SDS at pH 7.0.

scholarly journals THE METABOLIC CHARACTERISTICS OF NUCLEOLAR, CHROMOSOMAL, AND CYTOPLASMIC RIBONUCLEIC ACID OF DROSOPHILA SALIVARY GLANDS

1960 ◽  
Vol 8 (2) ◽  
pp. 365-378 ◽  
Author(s):  
Rachel McMaster-Kaye
Keyword(s):  
Salivary Glands ◽  
Ribonucleic Acid ◽  
Turnover Rate ◽  
Specific Activity ◽  
Rna Turnover ◽  
Metabolic Characteristics ◽  
Cytoplasmic Rna ◽  
Lower Maximum ◽  
Rna Accumulation ◽  
Nucleolar Rna

Incorporation and retention of adenine-8-C14 and of P32O4 by nucleolar, chromosomal, and cytoplasmic RNA have been studied. Radioisotope concentrations were determined from autoradiographs, by grain counting, and RNA concentrations by microphotometry after basic staining. The relation between rates of RNA accumulation and rates of adenine incorporation was used to determine if synthesis was used to replace RNA which was lost from a fraction, and to obtain estimates of turnover rate. Nucleolar incorporation patterns indicate its incorporation is independent of growth, and there is complete turnover of the fraction in an hour or less. Nucleolar turnover is attributed to degradation of RNA within the nucleolus rather than to movement of intact molecules from the nucleolus. Chromosomal RNA reaches a much lower maximum specific activity than nucleolar, and a slightly higher maximum than cytoplasmic RNA. It showed faster incorporation than cytoplasmic RNA while accumulating RNA at the same rate as the cytoplasm, suggesting chromosomal RNA turnover. No evidence of cytoplasmic RNA turnover was found: rate of incorporation and rate of growth were correlated, and retention studies detected no decrease in amount of RNA-C14, RNA-P32, or RNA. Different ultimate precursors are indicated for nucleolar and non-nucleolar RNA by the observation that the nucleolar precursor is labeled before the precursor of non-nucleolar RNA.

scholarly journals Plant mitochondrial pyruvate dehydrogenase complex: purification and identification of catalytic components in potato

1998 ◽  
Vol 334 (3) ◽  
pp. 571-576 ◽  
Author(s):  
A. Harvey MILLAR ◽  
Carina KNORPP ◽  
Christopher J. LEAVER ◽  
Steven A. HILL
Keyword(s):  
Pyruvate Dehydrogenase ◽  
Specific Activity ◽  
Divalent Cations ◽  
Pyruvate Dehydrogenase Complex ◽  
Protein Sequences ◽  
Protein Band ◽  
Sds Page ◽  
Molecular Masses ◽  
Α Subunits ◽  
Dehydrogenase Complex

The pyruvate dehydrogenase complex (mPDC) from potato (Solanum tuberosum cv. Romano) tuber mitochondria was purified 40-fold to a specific activity of 5.60 µmol/min per mg of protein. The activity of the complex depended on pyruvate, divalent cations, NAD+ and CoA and was competitively inhibited by both NADH and acetyl-CoA. SDS/PAGE revealed the complex consisted of seven polypeptide bands with apparent molecular masses of 78, 60, 58, 55, 43, 41 and 37 kDa. N-terminal sequencing revealed that the 78 kDa protein was dihydrolipoamide transacetylase (E2), the 58 kDa protein was dihydrolipoamide dehydrogenase (E3), the 43 and 41 kDa proteins were α subunits of pyruvate dehydrogenase, and the 37 kDa protein was the β subunit of pyruvate dehydrogenase. N-terminal sequencing of the 55 kDa protein band yielded two protein sequences: one was another E3; the other was similar to the sequence of E2 from plant and yeast sources but was distinctly different from the sequence of the 78 kDa protein. Incubation of the mPDC with [2-14C]pyruvate resulted in the acetylation of both the 78 and 55 kDa proteins.

scholarly journals Carbonic Anhydrase III Is Not Required in the Mouse for Normal Growth, Development, and Life Span

2004 ◽  
Vol 24 (22) ◽  
pp. 9942-9947 ◽  
Author(s):  
Geumsoo Kim ◽  
Tae-Hoon Lee ◽  
Petra Wetzel ◽  
Cornelia Geers ◽  
Mary Ann Robinson ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Carbon Dioxide ◽  
Carbonic Anhydrase ◽  
Physiological Function ◽  
Specific Activity ◽  
Normal Growth ◽  
Standard Laboratory ◽  
Wild Type ◽  
Carbonic Anhydrase Iii ◽  
Growth Development

ABSTRACT Carbonic anhydrase III is a cytosolic protein which is particularly abundant in skeletal muscle, adipocytes, and liver. The specific activity of this isozyme is quite low, suggesting that its physiological function is not that of hydrating carbon dioxide. To understand the cellular roles of carbonic anhydrase III, we inactivated the Car3 gene. Mice lacking carbonic anhydrase III were viable and fertile and had normal life spans. Carbonic anhydrase III has also been implicated in the response to oxidative stress. We found that mice lacking the protein had the same response to a hyperoxic challenge as did their wild-type siblings. No anatomic alterations were noted in the mice lacking carbonic anhydrase III. They had normal amounts and distribution of fat, despite the fact that carbonic anhydrase III constitutes about 30% of the soluble protein in adipocytes. We conclude that carbonic anhydrase III is dispensable for mice living under standard laboratory husbandry conditions.

scholarly journals Isolation of a human hepatic 60 kDa aspartylglucosaminidase consisting of three non-identical polypeptides

1989 ◽  
Vol 262 (1) ◽  
pp. 189-194 ◽  
Author(s):  
M Baumann ◽  
L Peltonen ◽  
P Aula ◽  
N Kalkkinen
Keyword(s):  
Gel Electrophoresis ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Polyacrylamide Gel ◽  
Inherited Disease ◽  
Ph Optimum ◽  
Native Polyacrylamide Gel Electrophoresis ◽  
Molecular Masses ◽  
Polypeptide Chains

We have characterized the properties of human aspartylglucosaminidase (EC 3.5.1.26), the lysosomal enzyme which is deficient in the human inherited disease aspartylglucosaminuria. The purification procedure from human liver included affinity chromatography, gel filtration, strong-anion- and strong-cation-exchange h.p.l.c., chromatofocusing and reverse-phase h.p.l.c. In a denaturing SDS/polyacrylamide-gel electrophoresis, the 6600-fold purified enzyme was shown to be composed of three non-identical inactive polypeptide chains of molecular masses 24, 18 and 17 kDa. In a native polyacrylamide-gel electrophoresis, these polypeptide chains ran as one active enzyme complex. As judged from the elution position of the native enzyme in a Biogel P-100 gel filtration, the approximate molecular mass of this complex was 60 kDa. The enzyme had a pI of 5.7, a pH optimum at 6, of 0.48 mM and a specific activity of 200,000 nkat for the substrate 2-acetamido-1-beta-(L-aspartamido)-1,2-dideoxy-D-glucose. The enzyme showed a 57% loss of activity at 60 degrees C after 45 h but was practically inactive after incubation at 72 degrees C for a few minutes. The molecular structure, Km and specific activity as well as the thermostability of the enzyme described here are different from those reported previously for human aspartylglucosaminidase.

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