Ribonucleic acid isolation

Glisin, V., Crkvenjakov, R, and Byus, C (1974) Ribonucleic acid isolation by cesium chlonde centrifugation. Biochemistry 13,2633-2637. 

N18 Nievel, J. G. Transport and incorporation of labelled orotate into ribonucleic acid isolated rat liver perfused with actinomycin D. Biochem. Soc. Trans., 3, 1239-1241 (1975)... 

Colter, J. S., Bird, H. H., Moyer, A. W., Brown, R. A. Infectivity of ribonucleic acid isolated from virus infected tissues. Virology 4, 522 (1957). 

Cytosine was isolated from hydrolysis of calf thymus in 1894 and by 1903 its structure was known and it had been synthesized from 2-ethylthiopyrimidin-4(3H)-one. The acid hydrolysis of ribonucleic acid gives nucleotides, among which are two cytidylic acids, 2 -and 3 -phosphates of cytidine further hydrolysis gives cytidine itself, i.e. the 1-/3-D-ribofuranoside of cytosine, and thence cytosine. The deoxyribonucleic acids likewise yield deoxyribonucleotides, including cytosine deoxyribose-5 -phosphate, from which the phosphate may be removed to give cytosine deoxyriboside and thence cytosine. 

Among the aldopentoses, D-ribose is a component of many biologically important substances, most notably the ribonucleic acids, and D-xylose is very abundant and is isolated by hydrolysis of the polysaccharides present in corncobs and the wood of trees. 

As is well-known, nucleic acids consist of a polymeric chain of monotonously reiterating molecules of phosphoric acid and a sugar. In ribonucleic acid, the sugar component is represented by n-ribose, in deoxyribonucleic acid by D-2-deoxyribose. To this chain pyrimidine and purine derivatives are bound at the sugar moieties, these derivatives being conventionally, even if inaccurately, termed as pyrimidine and purine bases. The bases in question are uracil (in ribonucleic acids) or thymine (in deoxyribonucleic acids), cytosine, adenine, guanine, in some cases 5-methylcytosine and 5-hydroxymethylcyto-sine. In addition to these, a number of the so-called odd bases occurring in small amounts in some ribonucleic acid fractions have been isolated. 

Gillam, I., Blew, D., Warrington, R. C., von Tigerstrom, M., and Tener, G. M. (1968). A general procedure for the isolation of specific transfer ribonucleic acids. Biochemistry 7,... 

A detailed discussion of the modes of occurrence and biological importance of the polynucleotides is outside the scope of this article. However, in examining the structures of polynucleotides, it is necessary to take into consideration the origins of the materials studied. The pioneer researches of Caspersson114 indicated that deoxyribonucleic acids are present exclusively in the nucleus, whereas ribonucleic acids are found chiefly in the cytoplasm and only to a small extent in the nucleus. This general outline of the distribution of nucleic acids within the cell has been confirmed and extended by more recent work,116 and it has been possible to isolate both types of nucleic acid from different cellular fractions of the same tissue.116... 

In a number of methods, isolation of the nucleoprotein complex (stage 2) is avoided. In the isolation of ribonucleic acid from beef pancreas,1241 nuclear material and cell debris are removed from a normal-saline extract of the minced tissue, which is then brought to half-saturation with sodium chloride (to dissociate the protein from the nucleic acid). After removal of the protein, the nucleic acid is precipitated with alcohol. However, the suggestion has been made126 that it is more satisfactory to isolate the nucleoprotein first, and this has been carried out, for instance, in the extraction of the ribonucleic acid from fowl sarcoma GRCH 15.126 Nucleoprotein complexes have also been isolated from baker s yeast127 and have been separated into various fractions, the nucleic acids from which differ slightly in composition. In addition, nucleoproteins have been isolated by complex formation with cetyltrimethylammonium bromide.128... 

The position was somewhat clarified by the isolation of 2- and 3-O-phos-phonucleosides from ribonucleic acid hydrolyzates in 92 to 100% yields,134 and also by the demonstration that 5-O-phosphonucleosides are also present in enzymic digests.49, 197 Yet this information gave no indication of the nature of the alkali-labile linkages. Thus, while the majority of the experimental evidence pointed to the phosphoryl residues as being doubly esterified with adjacent nucleosides, two facts remained apparently inexplicable on the basis of this type of structure. First, ready fission by alkalis, and secondly, the absence of 5-phosphates from alkaline hydrolyzates and their presence in enzymic digests. Both these facts have been explained by Brown and Todd in the following way.92... 

The C-nucleosides are a group of C-glycosylated heterocycles in which the anomeric carbon atom is attached to the heterocycle by a C-C bond. For a number of years after its discovery, pseudouridine1 (1) was the only representative of this class of compound it is found as a minor component in various transfer ribonucleic acids.2 Since 1959, a number of other C-nucleosides have been isolated in rapid succession, mainly from fermentation sources, and have been found to exhibit a variety of interesting biological properties.3 Thus, pyraz-... 

Chirgwin JM, Pryzbyla AE, MacDonald RJ, Rutter WJ. 1979. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18 5294-5299. 

Bismuth hydroxide is used as an absorbent and in the hydrolysis of ribonucleic acid. It also is used in the isolation of plutonium from irradiated uranium. 

The most important pyrimidine derivatives are those upon which biological organisms depend. Cytosine 1018 and uracil 1019 are found in ribonucleic acid (RNA) in the form of their ribonucleotides, cytidine 1020 and uridine 1021, while in deoxyribonucleic acid (DNA), cytosine and thymine 1022 are found in the form of their 2 -deoxyribonucleotides, 2 -deoxycytidine 1023 and thymidine 1024. 5-Methylcytosine 1025 is also found to a small extent (c. 5%) in human DNA in the form of its 2 -deoxyriboside 1026, and 5-(hydroxymethyl)cytosine-2 -deoxyriboside 1027 has also been detected in smaller amounts <2005CBI1>. Many variants of cytosine and uracil can be found in RNA including orotic acid 1028 in the form of its ribonucleotide orotidine 1029. Other pyrimidine derivatives to have been isolated from various biological sources include 2 -deoxyuridine 1030, alloxan 1031, and toxopyrimidine (pyramine) 1032 (Figure 2). 

Berger, S.L., and C.S. Birkenmeier. 1979. Inhibition of intractable nucleases with ribonucleoside-vanadyl complexes Isolation of messenger ribonucleic acid from resting lymphocytes. Biochemistry 18 5143. 

The term nucleoside was originally proposed by Levene and Jacobs in 1909 for the carbohydrate derivatives of purines (and, later, of pyrimidines) isolated from the alkaline hydrolyzates of yeast nucleic acid. The phosphate esters of nucleosides are the nucleotides, which, in polymerized forms, constitute the nucleic acids of all cells.2 The sugar moieties of nucleosides derived from the nucleic acids have been shown, thus far, to be either D-ribose or 2-deoxy-D-eri/fAro-pentose ( 2-deoxy-D-ribose ). The ribo-nucleosides are constituents of ribonucleic acids, which occur mainly in the cell cytoplasm whereas 2-deoxyribo -nucleosides are components of deoxypentonucleic acids, which are localized in the cell nucleus.3 The nucleic acids are not limited (in occurrence) to cellular components. They have also been found to be important constituents of plant and animal viruses. 

Not included in Figs. 1 and 2 are the several methylated-purine nucleosides found in certain nucleic acids. The deoxynucleoside of 6-methylaminopurine has been found10 in the deoxyribonucleic acid of Escherichia coli 15T- as well as in other microbial sources. Nucleosides of 6-methylaminopurine, 6-dimethylaminopurine, and 6-amino-2-methylpurine have also been isolated from the ribonucleic acid of Escherichia coli, as well as from other sources [J. W. Littlefield and D. B. Dunn, Nature, 181, 254 (1958)]. 

Since D-ribose derivatives of amines occur naturally in ribonucleic acids, vitamin B12, and in cozymase and are used in the isolation of D-ribose as well as in the synthesis of vitamin B2, they have attracted much attention and are currently under active investigation in a number of laboratories. As mentioned earlier, the ribonucleic acids and their component parts have been well reviewed elsewhere 4 in the following discussion the syntheses of the nucleosides will be discussed only insofar as they illustrate methods of forming N-ribosides. 

PolyP-ribonucleic acid complexes have been isolated from a variety of organisms (Belozersky, 1955,1958, 1959a Chayenetal. 1955 ChaloupkaandBabicky, 1957, 1958 ... 

D. L. Correll and N. E. Tolbert (1962). Ribonucleic acid-polyphosphate from algae. I. Isolation and... 

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