Ribonucleases of beef brain nuclei. Purification and characterization of an alkaline RNAase

Niedergang, Claude; Okazaki, H.; Ittel, Marie‐Elisabeth; Muñoz, D Martínez de; Petek, F; Mandel, P.

doi:10.1016/0005-2744(74)90261-7

Cited by 29 publications

(6 citation statements)

References 26 publications

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“…Alkaline ribonuclease activity was assayed as described by Niedergang et al (1974) including modifications introduced by Sajdel-Sulkowska and Marotta (1984). Frozen samples, taken from the same region of the neocortex used for each isolation of total cellular RNA, were homogenized in water and then adjusted to 60 mM citrate-phosphate; 0.5 mM EDTA, pH 7.5.…”

Section: Alkaline Rnase Assaymentioning

confidence: 99%

Characterization of Messenger RNA from the Cerebral Cortex of Control and Alzheimer‐Afflicted Brain

Guillemette

Wong

McLachlan

et al. 1986

Journal of Neurochemistry

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A detailed comparative study of RNA transcripts isolated from the neocortex of control and Alzheimer postmortem brains was made to determine whether morphological changes in the chromatin of Alzheimer neurons and glia, which we reported earlier, are accompanied by changes in the products of transcription. A number of parameters were determined including the yields of total and mRNA per gram of tissue, the relative proportions of polyadenylated [poly(A) +] mRNA in the total RNA, the size distribution of the transcripts and the length of their poly(A) tails, and the nature of their in vitro translation products. The levels of endogenous RNase activity were also measured. The effect of the agonal process on the transcript complement was examined by Northern blotting of a cloned human heat‐shock cDNA to total human brain RNA. Our results reveal that the yields of total RNA, unadenylated mRNA, and poly(A) tail lengths from Alzheimer neocortex samples do not differ significantly from those of control and non Alzheimer dementia neocortex. On the other hand we find a significant reduction in the levels and proportion of poly(A)+ mRNA in the Alzheimer samples as compared to control brain samples. Quantitative rather than qualitative differences were observed in the in vitro translation products when programmed with control and Alzheimer mRNA. No differences were found in the levels of RNase activity between control and Alzheimer simples. Heatshock mRNA transcripts were detected in brain samples from patients in whom fever was associated with death. The direct correlation of reduced poly(A)+ mRNA and chromatin condensation in Alzheimer neocortex suggests a cause‐and‐effect relationship. Whether all transcribed genes are affected or only a specific subset has yet to be determined.

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Section: Alkaline Rnase Assaymentioning

confidence: 99%

Characterization of Messenger RNA from the Cerebral Cortex of Control and Alzheimer‐Afflicted Brain

Guillemette

Wong

McLachlan

et al. 1986

Journal of Neurochemistry

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“…Comparison with Other Nucleases. The base specificity of this brain ribonuclease is similar to that of bovine pancreatic RNase (Figure 5), the major ribonuclease of bovine seminal plasma (Floridi et al, 1972), and probably the nuclease of beef brain nuclei (Niedergang et al, 1974), as well as the pancreatic RNases of other species. We are particularly interested in the relationship of the brain cnz.ymc to other bovine ribonucleases.…”

Section: Discussionmentioning

confidence: 56%

“…with acid and alkaline pH optima are present in many mammalian tissues including brain (Barnard, 1969;Datta et al, 1964;Roth and Milstein, 1952), and efforts have been directed at enzyme localization and purification (e.g., Aksenova and Nechaeva, 1971;Houck, 1958;Nechaeva, 1972;Niedergang et al, 1974).…”

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confidence: 99%

Characterization of a ribonuclease from bovine brain

Elson¹,

Glitz²

1975

Biochemistry

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An alkaline ribonuclease (pH optimum near 8) has been purified from whole beef brains and found to have a base specificity like that of bovine pancreatic ribonuclease, but in most other respects to be distinguishable from the enzymes of bovine pancreas, semen, or brain nuclei. The preparation appears homogeneous in sedimentation equilibrium and probably so in polyacrylamide gel electrophoresis under normal or dissociating conditions. Sedimentation equilibrium and SDS gel electrophoresis both indicate a molecular weight of 2.4-2.6 times 10-4, and tryptic and chymotrypic peptide patterns are consistent with a protein of this size. No dissociation into subunits has been attained. The enzyme is not precipitated by antiserum to pancreatic ribonuclease, although its activity is inhibited by this antiserum with low efficiency. In comparisons of the hydrolysis of RNA the brain enzyme was found to have a similar specificity to pancreatic RNase, but to have a loser Km for RNA and to produce significantly different oligonucleotides upon partial hydrolysis of bacteriophage RNA, suggesting differences in the mechanism of substrate recognition. In contrast, nuclease inactivation by iodoacetate at pH 5.5 is indistinguishable for pancreatic or purified brain RNase.

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“…The enzyme isolated is specific for poly(U) and poly(C) with higher affinity for poly(C) in comparison to poly(U). Isolation of similar RNases specific for poly(U) and poly(C), with higher rates for one of the two substrates, has been earlier reported for several tissues, e. g. beef brain cytosol, which is active against poly(C) but not against poly(U) [21]; bovine brain nuclei, which cleaves poly(U) at 20% of the rate of poly(C) [22] ; and bovine aorta, which attacks poly(C) at seven times the rate for poly(U) [23]. We believe that our enzyme is different from pancreatic ribonuclease and also from the other ribonucleases which show the same substrate specificity, for the following reasons.…”

Section: Discussionmentioning

confidence: 71%

Purification and characterization of a ribonuclease specific for poly(U) and poly(C) from the larvae of Ceratitis capitata

Sideris¹,

Fragoulis²

1987

European Journal of Biochemistry

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A specific ribonuclease was detected and purified to homogeneity from six-day-old larvae of the insect Ceratitis capitata and its homogeneity was checked by analysis in polyacrylamide gels in the presence of sodium dodecyl sulfate.The nuclease specifically degrades poly(U) and poly(C) whilst it fails to do so with other single-stranded homopolyribonucleotides. The enzyme has a pH optimum in the region 7 -9 and relative molecular mass of about 25 000.The effect of this ribonuclease on the integrity of RNAs isolated from six-day-old larvae or rat liver was also studied.The control of protein synthesis in eucaryotes is a fundamental problem not well understood. The fact that different mRNAs may be translated at different rates in eucaryotic cells has been firmly established. However, the molecular mechanism which determines the translation of an individual mRNA has not been yet fully understood. Recently researchers have adopted a new approach in order to solve the above problem and this is based on enzymes degrading RNA with special emphasis on endoribonucleases derived from different tissues and organisms. Some of the enzymes studied are a calf thymus endoribonuclease V, which specifically cleaves poly(A) and poly(U) [l], an endoribonuclease VII, which specifically degrades poly(U) and poly(C) [2], a chick oviduct ribonuclease IV, which specifically cleaves poly(A) [3], and an endoribonuclease specific for poly(U), isolated either from Artemia snlina [4] or patients with systemic lupus erythematosus [5].In this report we describe the isolation and purification to homogeneity of a ribonuclease derived from six-day-old larvae of Ceratitis capitata, which degrades poly(U) and poly(C). MATERIALS AND METHODS AnimaEsCeratitis capitata larvae were reared in a medium containing sucrose, yeast and cholesterol at 27°C and 75% relative Correspondence to

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Ribonucleases of beef brain nuclei. Purification and characterization of an alkaline RNAase

Cited by 29 publications

References 26 publications

Characterization of Messenger RNA from the Cerebral Cortex of Control and Alzheimer‐Afflicted Brain

Characterization of Messenger RNA from the Cerebral Cortex of Control and Alzheimer‐Afflicted Brain

Characterization of a ribonuclease from bovine brain

Purification and characterization of a ribonuclease specific for poly(U) and poly(C) from the larvae of Ceratitis capitata

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