ENRICHMENT OF SERINE-ACCEPTOR SOLUBLE RNA BY NUCLEIC ACID GELS.
Nucleic acid gels prepared by ultraviolet irradiation of polyuridylic acid on plastic beads can absorb a species of soluble RNA with increased serine-acceptor activity.
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Nucleic acid gels prepared by ultraviolet irradiation of polyuridylic acid on plastic beads can absorb a species of soluble RNA with increased serine-acceptor activity.
We have isolated a compound responsible for the cytokinin activity of soluble RNA from Escherichia coli. The structure, indicated as 6-(3-methyl-2-butenylamino)-2-methylthio-9-beta-D-ribofuranosylpurine, C(16)H(23)N(5)0(4)S, on the basis of low-and high-reso!ution mass spectrometry, was established by unequivocal synthesis. The mass spectra, chromatographic behavior, and ultraviolet spectra of the compounds from natural and synthetic sources were identical.
Kinin requiring tobacco and soybean tissues incubated on a medium containing N,6-benzyladenine-8-C(14) incorporated C(14) into several RNA components including adenylic and guanylic acids. About 15% of the label taken up by the tissues appeared in RNA while the remainder was distributed among several metabolites in the soluble, nonpolynucleotide fraction. Tissue grown on a kinin labeled in the side chain (N,6-benzyladenine-benzyl-C(14)) also incorporated a small, but nevertheless repeatable, amount of radioactivity into minor RNA components.Ultracentrifugation studies and methylated albumin chromatography indicated that the bulk of the label from benzyladenine-benzyl-C(14) is in soluble RNA. Approximately 50% of the C(14) in soluble RNA is in a component which has chromatographic properties like that of benzyladenine.It is suggested that the biological action of the kinins may hinge on their providing substituted bases in RNA in tissues which through differentiation no longer synthesize RNA-methylating enzymes. As an alternative it was hypothesized that a small amount of benzyladenine was incorporated into a m-RNA, acting there as a derepressing agent, perhaps by preventing its normal repressing function.
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Quantification of human immunodeficiency virus (HIV) RNA by branched-chain DNA signal amplification, measurement of soluble tumor necrosis factor type II receptors (sTNFR-II), neopterin, beta2-microglobulin, or CD4 cell counts can be used to predict the risk of clinical progression or death in HIV infection but have not been compared in the same study. Ninety subjects were categorized into progression groups by their rate of CD4 cell decline and matched into triplets by initial CD4 cell count, age, race, and calendar time. By matched logistic regression, only the sTNFR-II and HIV RNA values were predictive of outcome across the progression groups. Categorization of baseline HIV RNA and sTNFR-II resulted in differences in progression to several clinical outcomes. sTNFR-II concentrations were the only immune marker examined that increased the prognostic utility of HIV RNA determination in early-stage subjects. Further studies in later stages of disease or after therapy are indicated.
RNA polymerase enzymes isolated from soybean hypocotyl tissue during successive developmental stages (2-8 days old) have been fractionated by Sephadex column isoelectric focusing. Both the enzymes bound to chromatin and those enzymes free in the soluble phase were investigated during development with respect to their distribution within these two pools. All observed activites were classified according to their alpha-amanitin sensitivity and isoelectric points. Two Class I subspecies (Ia, Ib) and two Class III subspecies (IIIa, IIIb) were continually present bound to chromatin throughout the developmental sequences except the IIb form which was absent at the latest stage. However, a great multiplicity (9 total) of Class II activities (totally inhibited by alpha-amanitin) were observed to be bound to chromatin at the 2nd day stage. These forms were first released from the chromatin complex and recovered in a soluble pool (4th day stage). Subsequent hypocotyl development was accompanied by the gradual disappearance of these Class II subspecies from this pool (6th day) until only two soluble species and one chromatin-bound Class II activity remained (8th day). These observations indicate that the early development of this tissue is accompanied by a dramatic alteration in the conplexity of chromatin-bound RNA polymerase subspecies. Such events may in part determine the domain of RNA secies synthesized at successive developmental stages.
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