Purification, properties and specificity of an endonuclease from Agropyron elongatum seedlings

Yupsanis, Traianos; Symeonidis, L.; Kalemi, Theodora G.; Moustaka, H.; Yupsani, Anastasia

doi:10.1016/j.plaphy.2004.09.002

Cited by 19 publications

(3 citation statements)

References 37 publications

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“…After homogenization, the mixture was centrifuged at 24,000 g at 4°C for 15 min, the supernatant containing the released nucleotides was recovered, and its absorbance was measured at 260 nm. One unit of nuclease is defined as the amount of enzyme that produce a ∆OD 260 = 0.01 min −1 (Yupsanis et al, 2004).…”

Section: Determination Of Nuclease Activitymentioning

confidence: 99%

Nucleoside Metabolism Is Induced in Common Bean During Early Seedling Development

et al. 2021

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Nucleoside hydrolases (NSH; nucleosidases) catalyze the cleavage of nucleosides into ribose and free nucleobases. These enzymes have been postulated as key elements controlling the ratio between nucleotide salvage and degradation. Moreover, they play a pivotal role in ureidic legumes by providing the substrate for the synthesis of ureides. Furthermore, nucleotide metabolism has a crucial role during germination and early seedling development, since the developing seedlings require high amount of nucleotide simultaneously to the mobilization of nutrient in cotyledons. In this study, we have cloned two nucleosidases genes from Phaseolus vulgaris, PvNSH1 and PvNSH2, expressed them as recombinant proteins, and characterized their catalytic activities. Both enzymes showed a broad range of substrate affinity; however, PvNSH1 exhibited the highest activity with uridine, followed by xanthosine, whereas PvNSH2 hydrolyses preferentially xanthosine and shows low activity with uridine. The study of the regulation of nucleosidases during germination and early postgerminative development indicated that nucleosidases are induced in cotyledons and embryonic axes just after the radicle emergence, coincident with the induction of nucleases activity and the synthesis of ureides in the embryonic axes, with no remarkable differences in the level of expression of both nucleosidase genes. In addition, nucleosides and nucleobase levels were determined as well in cotyledons and embryonic axes. Our results suggest that PvNSH1 and PvNSH2 play an important role in the mobilization of nutrients during this crucial stage of plant development.

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Section: Determination Of Nuclease Activitymentioning

confidence: 99%

Nucleoside Metabolism Is Induced in Common Bean During Early Seedling Development

et al. 2021

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“…Additionally, the presence of protein band with molecular weight of 57.8 might be cyclin dependent kinase inhibitors that have been formerly found in plants and called putative CKIs as reported by Jasinski et al (2002). In addition, proteins with molecular mass of 18, 29, 31, 35 and 51KDa could be considered as protein bands for several types of nucleases enzymes that are responsible for DNA degradation as was reported by Yupsanis et al (2004).…”

Section: Discussionmentioning

confidence: 85%

Assessment of antimitotic and programmed cell death stimulation potentials of Galium sinaicum (Delile ex Decne) Boiss. at cellular level

Sobieh

2021

Caryologia

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Galium sinaicum is a wild medicinal plant in saint Catherine, Egypt. To distinguish apoptotic effect of G. sinaicum ethanol extract (GsEE), we examined the role of GsEE in inducing programmed cell death (PCD) of Allium cepa root meristematic cell (AcR). Cells was subjected to GsEE in definite concentrations (0.1,0.3, 0.5%) and duration (6, 12h), then PCD induction was assessed. Application of GsEE arrested the mitotic division of AcR with metaphase accumulation. Electron microscopy analysis demonstrated ultrastructural alterations of organelles verifying PCD hallmarks. Protein electrophoresis analysis of AcR revealed a change in protein profile of Allium cepa root, also quantitative analysis showed significant increase in nuclease activity enzymes that stimulated DNA laddering fragmentation. Additionally, cell proliferation of MCF-7 and BHK21 was arrested by GsEE. Apoptotic effect of G. sinaicum may be attributed to the presence of potent phenolic compounds such as querectin and rutin as established by HPLC phenolic fingerprint analysis.

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“…These enzymes are non-specific toward the sugar residue of the substrate and degrade both single-stranded DNA (ssDNA) and RNA. Although the individual functions of plant nucleases in vivo are not well defined, it is considered that they participate in nucleic acids degradation during programmed cell death (Aoyagi et al 1998;Thomas et al 2003) and have central role in DNA replication, recombination, and repair in higher plants (Grafi and Larkins 1995;Sugiyama et al 2000;Yupsanis et al 2004). According to pH optimum plant, nucleases are distinguished to acid and neutral.…”

Section: Introductionmentioning

confidence: 99%

Paraquat and roundup effects on nucleases activities of alfalfa seedlings and alfalfa nucleases activities on paraquat-treated and roundup-treated nucleic acids

et al. 2009

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The specific activities of acid (pH 5.5) and neutral (pH 7) DNases and RNases were determined in alfalfa (Medicago sativa L.) seedlings grown in the dark in the presence of 3.7 mM paraquat (PQ) or 1 mM roundup (RD). Seedlings were taken at 0, 1, 3, and 5 days. Plant growth parameters (plant height and fresh weight) were dramatically reduced under these conditions of growth comparing to the control (grown in water). The DNase and RNase specific activities of herbicide-treated seedlings were reduced. The reduction of activities ranged by about 50-90 and 15-70% in PQ-and RD-treated seedlings, respectively. In vitro, PQ-and RD-treated nucleic acids [single-stranded DNA (ssDNA), RNA, and plasmid DNA (pl-DNA)] were incubated with acid and neutral nucleases. Both enzymes were isolated and purified from alfalfa seedlings. Electrophoretic analysis on agarose gel of the above incubated mixtures revealed the following: (a) neutral nuclease (pH 7) was capable of hydrolyzing PQ-treated ssDNA while acid nuclease (pH 5.5) was incapable. This could be due to the fact that acid and neutral nucleases displayed different base linkage specificity toward ssDNA; (b) RD formed strong complexes with ssDNA that were unable to be hydrolyzed by both nucleases; (c) in contrast, both enzymes were capable of hydrolyzing PQ-or RD-treated RNA; (d) neutral nuclease was capable of nicking and linearizing both PQ-and RDtreated pl-DNA while acid nuclease had the same activity only toward the PQ-treated pl-DNA; (e) the enzyme activities were not inhibited in the presence of both herbicides. The data suggest that the complexes of PQ or RD with DNA should not be functional substrates of nucleases, and consequently cell processes (e.g., metabolism of nucleic acids, gene expression, replication), in which DNA and nucleases are involved, could be disturbed.

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Purification, properties and specificity of an endonuclease from Agropyron elongatum seedlings

Cited by 19 publications

References 37 publications

Nucleoside Metabolism Is Induced in Common Bean During Early Seedling Development

Nucleoside Metabolism Is Induced in Common Bean During Early Seedling Development

Assessment of antimitotic and programmed cell death stimulation potentials of Galium sinaicum (Delile ex Decne) Boiss. at cellular level

Paraquat and roundup effects on nucleases activities of alfalfa seedlings and alfalfa nucleases activities on paraquat-treated and roundup-treated nucleic acids

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