scholarly journals Ribosomal and informational ribonucleoprotein complexes of animal cells. Study on rat liver ribonucleic acids as constituents of ribonucleoprotein complexes by chromatography on nucleoprotein-celite columns.

1975 ◽  
Vol 147 (3) ◽  
pp. 447-456 ◽  
Author(s):  
A V Lichtenstein ◽  
R P Alechina ◽  
V S Shapot

A novel method of RNA fractionation has been developed. Nuclear and cytoplasmic rat liver RNA species were fractionated as constituents of corresponding ribonucleoprotein particles, which were previously adsorbed on a Celite-column by their protein component. The fractionation is based on a dissociation of the particles (linear concentration gradient of LiCl and urea with subsequent temperature gradient), which results in a gradual release of the RNA molecules from ribonucleoprotein complexes. Thus the fractionation is in accordance with the tightness of the RNA-protein bonds. A gradient elution of RNA from a nucleoprotein-Celite column permitted fractionation of both ribosomal and rapidly labelled non-ribosomal RNA. The latter, both nuclear and cytoplasmic, could be separated by chromatography on nucleoprotein-Celite columns into two main fractions (components I and V). In cytoplasmic RNA components I and V presumably correspond to mlRNA (messenger-like RNA of free cytoplasmic particles) and mRNA (template RNA associated with ribosomes) respectively.

1971 ◽  
Vol 125 (1) ◽  
pp. 225-234 ◽  
Author(s):  
A. V. Lichtenstein ◽  
V. S. Shapot

1. The conformation of RNA was found to affect its behaviour on methylated albumin–kieselguhr chromatography. The less regular the secondary structure of RNA, the more tightly it binds to the methylated albumin–kieselguhr column. 2. The presence of various denaturing agents (such as urea or perchlorate) in the medium while RNA was adsorbed on the column increased the resolving power of the technique as exemplified by the separation of rat liver rRNA into two distinct peaks. A special procedure for selective adsorption of the cytoplasmic DNA-like RNA on the preparative scale has been developed. Polyribosomal mRNA (rapidly labelled RNA formed in the presence of small doses of actinomycin D) can also be adsorbed selectively by the column. 3. A type of tissue specificity was detected in nuclear RNA from rat liver, kidney, thymus and spleen by using a modified salt and temperature gradient for the chromatographic fractionation (Lichtenstein, Piker & Shapot, 1967; Shapot, Lichtenstein & Piker, 1967). It was also found that cytoplasmic RNA from the different rat tissues contained no tenaciously bound fraction at all, whereas it constituted about 50% of the nuclear RNA. The problem of the possible biological function of the tenaciously bound fraction is discussed.


2020 ◽  
Vol 17 ◽  
Author(s):  
LiJuan Wang ◽  
Yan Liu ◽  
Rui Li ◽  
DongXian He

Objectives: Triptolide (TPL) has been shown to have a good clinical effect on rheumatoid arthritis (RA). We designed TPL microspheres (TPL-MS) and investigated its metabolic behavior in human, dog, rabbit and rat liver microsomes (HLM, DLM, RLM and SDRLM) with UPLC-MS/MS method. Methods: First, a UPLC-MS/MS method was established to measure concentration of TPL in samples. The sample was separated on a C18 column (2.1×100 mm, 1.8μm) and eluted with a gradient elution. The precursor ion/product ion were m/z 378.1/361.0 for TPL and 260.0/116.2 for the internal standard. Then T1/2, Vmax and CLint were calculated from the above data. Finally, the metabolites of TPL-MS were identified by high-resolution UPLC-MS/MS. The sample was separated on a C18 column (2.1×100 mm, 2.2 μm) and eluted with isocratic elution. Mass spectrometric detection was carried out on a thermo Q-exactive mass spectrometer with HESI. The scanning range of precursor ions was from m/z 50 to m/z 750. Result and Discussion: Through several indicators including standard curve, precision, accuracy, stability, matrix effect and recovery rate, the enzymatic kinetics parameters including T1/2, Vmax and CLint were completed. Several metabolites of TPL-MS were identified. Conclusion: UPLC-MS/MS method is an accurate and sensitive method for determination of TPL in liver microsome samples with good precision, accuracy and stability. The variation of parameters indicated that the microspheres can delay the elimination of TPL in liver microsomes. The metabolism of TPL-MS varied among species, but no new metabolites appeared.


eLife ◽  
2015 ◽  
Vol 4 ◽  
Author(s):  
André Plagens ◽  
Michael Daume ◽  
Julia Wiegel ◽  
Lennart Randau

Signal recognition particles (SRPs) are universal ribonucleoprotein complexes found in all three domains of life that direct the cellular traffic and secretion of proteins. These complexes consist of SRP proteins and a single, highly structured SRP RNA. Canonical SRP RNA genes have not been identified for some Thermoproteus species even though they contain SRP19 and SRP54 proteins. Here, we show that genome rearrangement events in Thermoproteus tenax created a permuted SRP RNA gene. The 5'- and 3'-termini of this SRP RNA are located close to a functionally important loop present in all known SRP RNAs. RNA-Seq analyses revealed that these termini are ligated together to generate circular SRP RNA molecules that can bind to SRP19 and SRP54. The circularization site is processed by the tRNA splicing endonuclease. This moonlighting activity of the tRNA splicing machinery permits the permutation of the SRP RNA and creates highly stable and functional circular RNA molecules.


1971 ◽  
Vol 26 (12) ◽  
pp. 1282-1287 ◽  
Author(s):  
P. G. Popov ◽  
L. I. Valeva-Dimitrova ◽  
A. A. Hadjiolov

Intact and adrenalectomized rats were irradiated with 900 R or treated with 10 mg per 100 g body weight of hydrocortisone. The incorporation of orotic acid-6-14C for 2 hours into liver free uridine nucleotides, nuclear and cytoplasmic RNA and RNA fractions obtained by agar gel electrophoresis, were studied.The obtained results show that in intact animals both irradiation and hydrocortisone induce a higher labelling of cytoplasmic and nuclear liver RNA. The higher labelling of RNA is not correlated with a higher labelling of the free uridine nucleotides. The labelling of all electrophoretic RNA fractions is increased to approximately the same extent under the action of either hydrocortisone or irradiation.Irradiation or hydrocortisone treatment of adrenalectomized rats causes also a higher labelling of liver nuclear and cytoplasmic RNA. The higher labelling of RNA is not correlated with that of free uridine nucleotides and affects all electrophoretic RNA fractions.. The combined action of irradiation and hydrocortisone shows an additive effect on the labelling of nuclear and cytoplasmic liver RNA.The obtained results indicate that whole body irradiation causes an increased synthesis of both ribosomal and non-ribosomal RNA’s of rat liver. Since the same effect is observed in intact and adrenalectomized animals it may be concluded that the stimulation of liver RNA synthesis by irradiation is not mediated by the adrenals.


1998 ◽  
Vol 30 (4) ◽  
pp. 252-258 ◽  
Author(s):  
T. Kondo ◽  
S. Okamoto ◽  
T. Todoroki ◽  
T. Hirano ◽  
F.W. Schildberg ◽  
...  

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