Isolation from Avena leaf tissues of a nuclease with the same type of specificity towards RNA and DNA accumulation of the enzyme during leaf senescence

Wyen, N.V.; Erdei, Sára; Farkas, G. L.

doi:10.1016/0005-2787(71)90601-0

Cited by 56 publications

(19 citation statements)

References 29 publications

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“…The increase in nuclease activity of leaf extracts is evident in NaDodSO4 activity gels as well as in solution assays, so (19). Moreover, we have found that barley seedlings held in the dark for 5 days exhibit a 31% loss of chlorophyll and a 3.6-fold increase in nuclease activity; DNA-containing NaDodSO4 activity gels show increased intensity of a band that migrates slightly faster than the major nuclease bands in wheat seedlings (unpublished data).…”

Section: Discussionmentioning

confidence: 72%

Single-strand-preferring nuclease activity in wheat leaves is increased in senescence and is negatively photoregulated

Blank

McKeon

1989

Proc. Natl. Acad. Sci. U.S.A.

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Single-strand-preferring nucleases (EC 3.1.30.1) selectively cleave internucleotide bonds in singlestranded regions ofpredominantly duplex DNA and DNARNA hybrids and extensively degrade denatured DNA and RNA.The functions of single-strand-preferring nuclease in plants are unknown. We have monitored this nuclease activity in flag leaves of wheat (Tritcum aestivum L. cv. Chinese Spring) undergoing natural senescence and in primary leaves of wheat seedlings undergoing dark-induced senescence. In flag leaves, nuclease activity remained at basal levels during the first 2 weeks after anthesis, while chlorophyll content increased to a maximum. Nuclease activity then rose in concert with a decline in chlorophyll, reaching a 16-fold elevation at 5 weeks postanthesis, when 53% of the chlorophyll had been lost. When 8-day-old wheat seedlings were induced to senesce by placing them in darkness, nuclease activity rose without apparent lag, reaching a 13-fold elevation in 7 days, when 61% of the chlorophyll had been lost. The increase in nuclease activity was reversible upon reexposure of seedlings to light, a decline beginning without apparent lag. Reversibility was complete for plants that had been held in darkness for 5 days, with activity returning to the control level in 2 days. These senescencerelated changes in nuclease activity, measured in conventional assays, were consistent with concomitant analysis by activity staining of sodium dodecyl sulfate/polyacrylamide gels. We conclude that an increase in single-strand-preferring nuclease activity is closely associated with wheat leafsenescence and that nuclease activity is subject to negative photoregulation.Endonucleases with a strong preference for polynucleotide substrates lacking ordered structure (EC 3.1.30.1) are well known in plants and in fungi (for reviews see refs. 1-4). These enzymes, such as S1 nuclease, discriminate between doubleand single-stranded polynucleotides and have thus become an important reagent for identification and selective hydrolysis of single-stranded regions of DNA and DNA-RNA hybrids. More is known, in fact, about research applications of these single-strand-preferring (SSP) nucleases than is known about their physiological functions. The ability to selectively cleave single-stranded DNA is well suited for biological processes that require precise and limited restructuring of predominantly duplex DNA. Indeed, a role for SSP nuclease in recombination and DNA repair in fungi is indicated by the isolation of nuclease-deficient mutants that are recombination-defective and ultraviolet-sensitive (2). Selective cleavage of single-stranded regions of tRNA is also known to occur in vitro. On the other hand, the capacity of SSP nuclease to extensively degrade single-stranded polynucleotides (e.g., mRNA) is compatible with other, quite different biological roles. Despite its catalytic versatility, no specific functions have been documented for the nuclease in plants. This situation is especially interesting because plant tissues have been foun...

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Section: Discussionmentioning

confidence: 72%

Single-strand-preferring nuclease activity in wheat leaves is increased in senescence and is negatively photoregulated

Blank

McKeon

1989

Proc. Natl. Acad. Sci. U.S.A.

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“…Their presence has also been shown in the endoplasmic reticulum, Golgi apparatus, protein bodies and vacuoles of the aleurone layer of barley seeds [38], bound to chromatin in the embryo axis of germinating pea [39], cultured tobacco cells [40] and leaves of Avena [41], spinach [42] and tea [43]. Moreover, they have also been isolated from mung bean sprouts [44], germinating pea seeds [45] and barley [46], potato tubers [47] and tobacco pollen [48].…”

Section: Occurrence and Localizationmentioning

confidence: 99%

Single-strand-specific nucleases

Desai

2003

FEMS Microbiology Reviews

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Single-strand-specific nucleases are multifunctional enzymes and widespread in distribution. Their ability to act selectively on singlestranded nucleic acids and single-stranded regions in double-stranded nucleic acids has led to their extensive application as probes for the structural determination of nucleic acids. Intracellularly, they have been implicated in recombination, repair and replication, whereas extracellular enzymes have a role in nutrition. Although more than 30 single-strand-specific nucleases from various sources have been isolated till now, only a few enzymes (S1 nuclease from Aspergillus oryzae, P1 nuclease from Penicillium citrinum and nucleases from Alteromonas espejiana, Neurospora crassa, Ustilago maydis and mung bean) have been characterized to a significant extent. Recently, some of these enzymes have been cloned, their crystal structures solved and their interactions with different substrates have been established. The detection, purification, characteristics, structure^function correlations, biological role and applications of single-strand-specific nucleases are reviewed.

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“…In terms of the evolutional change of plant RNases, knowledge is limited to those plants belonging to Dicotyledoneae, except for barley. 10) These include several plants of Solanaceae [tomato, 4,11) and tobacco, 3,12) petunia 13) and a kind of potato 14) ], Cucurbitaceae [bitter gourd, 7) cucumber, 8) melon 9) ], Rosacease [pear 15) ], Arabidopsis, 16) Zinnia elegance, 17) Panax ginnseng 18) and others. In this report, we will describe the amino acid sequence of an RNase from rice bran (Oryza sativa) which is a member of Monocotyledoneae, and its enzymatic properties.…”

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confidence: 99%

Amino Acid Sequence and Characterization of a Rice Bran Ribonuclease

Iwama

Ogawa

Yamagishi

et al. 2001

Biological & Pharmaceutical Bulletin

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RNase T2 family enzymes which are base non-specific and have a molecular mass of about 24 kDa, distributed widely from viruses to microbes, plants and animals. 1)They share two well conserved active site sequences, CAS I and CAS II. Therefore, they are a good model with which to study the evolution of RNases. In the plant kingdom, members of the RNase T2 family are classified into two classes, S-RNases which are the products of S-genes and function as self-incompatibility factors, 2) and S-like RNases.3) The latter enzymes are known to be induced by phosphate deprivation [Jost et al.,4) Bariola et al. 5)], in the case of senescence [Lers et al. 6) ], and exist in the seeds probably for defense [Ide et al.,7) Rojo et al. 8,9) ]. In terms of the evolutional change of plant RNases, knowledge is limited to those plants belonging to Dicotyledoneae, except for barley. 10) These include several plants of Solanaceae [tomato,4,11) and tobacco, 3,12) petunia 13) and a kind of potato 14) ], Cucurbitaceae [bitter gourd, 7) cucumber, 18) and others. In this report, we will describe the amino acid sequence of an RNase from rice bran (Oryza sativa) which is a member of Monocotyledoneae, and its enzymatic properties. MATERIALS AND METHODSEnzymes Lysylendopeptidase and g-pyroglutamine aminopeptidase were obtained from Wako Pure Chemical Ind., Ltd. (Osaka, Japan) and Calbiochem-Novabiochem. Co., Ltd., (LaJolla, LA, U.S.A.), respectively.Other Reagents Yeast RNA was a product of Marin Bio (Tokyo, Japan). Sixteen dinucleoside phosphates were purchased from Sigma (St. Louis, MO, U.S.A.). Phospho-cellulose and DE-52 were obtained from Whatman (Clifton, NJ, U.S.A.). A Mono Q column was obtained from Amersham Pharmcia Biotech (Buckinghamshire, England).Enzyme Assay (i) RNA as substrate: RNase assay was carried out as described 19) with RNA as substrate at pH 5.5 and 37°C. (ii) Dinucleosidephosphates as substrate: The rates of hydrolysis of 16 dinucleoside phosphates (XpYs), where X and Y were one of A, G, C and U, were measured spectrophotometrically as described by Imazawa et al. 20) in 0.1 M sodium acetate buffer (pH 5.5) at 22°C.Protein Concentration The protein concentration was measured spectrophotometrically assuming the absorbancy at 280 nm of a 0.1% protein solution to be 1.0.Purification of an RNase from Rice Bran Purification of an RNase from rice bran was performed by modification of the procedure reported by Yokoyama et al. 21)Step 1. Four kilograms of rice bran (product from a strain of Oryza sativa, Koshihikari) was suspended in 12 l of 20 mM sodium phosphate buffer (pH 7.0) and extracted by vigorous stirring at 4°C. The extract was filtered through a layer of cotton sheet, and the filtrate was centrifuged at 10000 rpm for 10 min. The pH of the supernatant was adjusted to 5.0 by addition of dil. HCl. The supernatant was kept at 4°C overnight, and the precipitate formed was eliminated by centrifugation at 10000 rpm for 10 min (crude extract).Step 2. The crude extract was fractionated by ammonium sulfate, and the pr...

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Isolation from Avena leaf tissues of a nuclease with the same type of specificity towards RNA and DNA accumulation of the enzyme during leaf senescence

Cited by 56 publications

References 29 publications

Single-strand-preferring nuclease activity in wheat leaves is increased in senescence and is negatively photoregulated

Single-strand-preferring nuclease activity in wheat leaves is increased in senescence and is negatively photoregulated

Single-strand-specific nucleases

Amino Acid Sequence and Characterization of a Rice Bran Ribonuclease

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