scholarly journals The proteins of Xenopus ovary ribosomes

1971 ◽  
Vol 125 (4) ◽  
pp. 1091-1107 ◽  
Author(s):  
P J Ford
Keyword(s):  
Potassium Chloride ◽  
Ribosomal Proteins ◽  
Large Subunit ◽  
Buoyant Density ◽  
Small Subunit ◽  
Chemical Methods ◽  
Ribosomal Subunits ◽  
Molecular Weights ◽  
Caesium Chloride ◽  
Chloride Treatment

1. The preparation of ribosomes and ribosomal subunits from Xenopus ovary is described. 2. The yield of once-washed ribosomes (buoyant density in caesium chloride 1.601g·cm-3; 44% RNA, 56% protein by chemical methods) was 10.1mg/g wet wt. of tissue. 3. Buoyant density in caesium chloride and RNA/protein ratios by chemical methods have been determined for ribosome subunits produced by 1.0mm-EDTA or 0.5m-potassium chloride treatment and also for EDTA subunits extracted with 0.5m-, 1.0m- or 1.5m-potassium chloride, 4. Analysis of ribosomal protein on acrylamide gels at pH4.5 in 6m-urea reveals 24 and 26 bands from small and large EDTA subunits respectively. The actual numbers of proteins are greater than this, as many bands are obviously doublets. 5. Analysis of the proteins in the potassium chloride extract and particle fractions showed that some bands are completely and some partially extracted. Taking partial extraction as an indication of possible doublet bands it was found that there were 12 and 20 such bands in the small and large subunits respectively, making totals of 36 and 46 proteins. 6. From the measured protein contents and assuming weight-average molecular weights for the proteins of large and small subunits close to those observed for eukaryote ribosomal proteins it is possible to compute the total numbers of protein molecules per particle. It appears that too few protein bands have been identified on acrylamide gels to account for all the protein in the large subunit, but probably enough for the small subunit.

scholarly journals Enzymic iodination of eukaryotic ribosomal subunits. Characterization and analysis by two-dimensional gel electrophoresis

1975 ◽  
Vol 152 (2) ◽  
pp. 373-378 ◽  
Author(s):  
David P. Leader
Keyword(s):  
Rat Liver ◽  
Gel Electrophoresis ◽  
Ribosomal Proteins ◽  
Large Subunit ◽  
Small Subunit ◽  
Suitable Method ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Ribosomal Subunits

1. Conditions are described for the enzymic iodination of ribosomal subunits from rat liver. The reaction is relatively insensitive to broad changes in the concentration of KCl, allowing subunits to be studied under conditions which minimize their dimerization. 2. Mixtures of extracted ribosomal proteins were iodinated with 125I, the proteins separated by two-dimensional gel electrophoresis and the radioactivity in each protein was determined. Thus 19 out of 23 of the proteins of the small subunit and 25 out of 33 of the proteins of the large subunit were labelled. Iodination should therefore be a suitable method for studying the topography of the ribosomal proteins of rat liver. 3. When the intact 40S subunit (rather than the extracted mixture of proteins) was iodinated, 18 of the 19 proteins were still labelled. However five of these were labelled less strongly than before. When the intact 60S subunit was iodinated, 17 of the 25 proteins were still labelled, although six of these were labelled less strongly. 4. These results show that in rat liver most of the ribosomal proteins of both subunits are at least partially at the surface of the particles. They are also consistent with the idea that the proportion of the ribosomal proteins in the interior of the particle may be greater for the 60S subunit than for the 40S subunit.

Separation of cytoplasmic ribosomal proteins of Microsporum canis

1987 ◽  
Vol 33 (4) ◽  
pp. 339-343 ◽  
Author(s):  
Valsan Mandiyan ◽  
G. Ramananda Rao
Keyword(s):  
Gel Electrophoresis ◽  
Ribosomal Proteins ◽  
Large Subunit ◽  
Polyacrylamide Gel Electrophoresis ◽  
Small Subunit ◽  
Polyacrylamide Gel ◽  
Microsporum Canis ◽  
Two Dimensional ◽  
Molecular Weights

The cytoplasmic ribosomal proteins of Microsporum canis were characterised in basic–acidic and basic–SDS two-dimensional polyacrylamide gel electrophoresis systems. The small subunit contained 28 proteins and the large subunit 38 proteins. The molecular weights of these proteins were in the range of 32 500 to 7600 and 48 000 to 11 000 in the small and large subunits, respectively. The 80S ribosomes showed 65 and 66 protein spots in basic–acidic and basic–SDS gel systems, respectively.

II. Mitt.: Auflösung der Chromoproteidfraktion aus Rattenleber-Ribosomen mit Hilfe der Polyacrylamidgel-Elektrophorese

1970 ◽  
Vol 25 (1) ◽  
pp. 67-72 ◽  
Author(s):  
Wolfram Domschke ◽  
Jürgen G. Meyer-Bertenrath
Keyword(s):  
Gel Electrophoresis ◽  
Ribosomal Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
The Other ◽  
Blue Staining ◽  
Ribosomal Subunits ◽  
Molecular Weights ◽  
The One ◽  
Liver Ribosomes

After preparation of a coloured protein component containing iron from rat liver ribosomes 1 this fraction was submitted to detailed analysis by means of polyacrylamide gel electrophoresis. Thus it may be separated into one main and two secondary bands, which do not contain RNA detectable by methylene blue staining. The ferric content of all bands can be demonstrated by staining with 2,4-dinitroso-1,3-naphthalenediol 2. These bands are to be found in the large ribosomal subunits as well as in the small one in qualitative conformity, they differ, however, in their quantitative relations to each other depending on origin. LiCl-extraction as described for the preparation of ribosomal proteins causes dissociation of the chromoproteid fraction into six bands possessing lower molecular weights each than the original bands.The chromoproteids, localized in the large ribosomal subunits on the one hand and in the small subunits on the other hand were prepared differentiatedly by gel filtration. Results show the chromoproteid represented in the 57-S-subunit on a bigger scale than the nucleoproteid part, on the contrary, the 29-S-subunit is constructed of RNA-containing material preferably.

Subcellular distribution of ribosomal proteins in Dictyostelium discoideum

1990 ◽  
Vol 68 (5) ◽  
pp. 839-845
Author(s):  
S. Ramagopal
Keyword(s):  
Ribosomal Proteins ◽  
Large Subunit ◽  
Small Subunit ◽  
Cellular Slime Mold ◽  
Developmentally Regulated ◽  
Two Dimensional Gel Electrophoresis ◽  
Coomassie Blue ◽  
Cellular Slime ◽  
State Growth

The distribution of ribosomal proteins in monosomes, polysomes, the postribosomal cytosol, and the nucleus was determined during steady-state growth in vegetative amoebae. A partitioning of previously reported cell-specific ribosomal proteins between monosomes and polysomes was observed. L18, one of the two unique proteins in amoeba ribosomes, was distributed equally among monosomes and polysomes. However S5, the other unique protein, was abundant in monosomes but barely visible in polysomes. Of the developmentally regulated proteins, D and S6 were detectable only in polysomes and S14 was more abundant in monosomes. The cystosol revealed no ribosomal proteins. On staining of the nuclear proteins with Coomassie blue, about 18, 7 from 40S subunit and 11 from 60S subunit, were identified as ribosomal proteins. By in vivo labeling of the proteins with [35S]methionine, 24 of the 34 small subunit proteins and 33 of the 42 large subunit proteins were localized in the nucleus. For the majority of the ribosomal proteins, the apparent relative stoichiometry was similar in nuclear preribosomal particles and in cytoplasmic ribosomes. However, in preribosomal particles the relative amount of four proteins (S11, S30, L7, and L10) was two- to four-fold higher and of eight proteins (S14, S15, S20, S34, L12, L27, L34, and L42) was two- to four-fold lower than that of cytoplasmic ribosomes.Key words: cellular slime mold, cell-specific ribosomal proteins, nucleus, cytoplasm, two-dimensional gel electrophoresis.

scholarly journals Particle weights and protein composition of the ribosomal subunits of the extremely thermoacidophilic archaebacterium Caldariella acidophila

1983 ◽  
Vol 209 (2) ◽  
pp. 461-470 ◽  
Author(s):  
P Londei ◽  
A Teichner ◽  
P Cammarano ◽  
M De Rosa ◽  
A Gambacorta
Keyword(s):  
Molecular Weight ◽  
Average Molecular Weight ◽  
Buoyant Density ◽  
Protein Composition ◽  
Equilibrium Density ◽  
Number Average Molecular Weight ◽  
Ribosomal Subunits ◽  
Molecular Weights ◽  
Protein Components ◽  
Protein Number

1. The ribosomal subunits of one thermoacidophilic archaebacterium (Caldariella acidophila) and of two reference eubacterial species (Bacillus acidocaldarius, Escherichia coli) were compared with respect to ribosome mass and protein composition by (i) equilibrium-density sedimentation of the particles in CsCl and (ii) gel-electrophoretic estimations of the molecular weights of the protein and the rRNA. 2. By either procedure, it is estimated that synthetically active archaebacterial 30S subunits (52% protein by wt.) are appreciably richer in protein than the corresponding eubacterial particles (31% protein by wt.) 3. The greater protein content of the archaebacterial 30S subunits is accounted for by both a larger number and a greater average molecular weight of the subunit proteins; specifically, C. acidophila 30S subunits yield 28 proteins whose combined mass is 0.6×10(6) Da, compared with 20 proteins totalling 0.35×10(6) Da mass for eubacterial 30S subunits. 4. No differences in protein number are detected among the large subunits, but C. acidophila 50S subunits exhibit a greater number-average molecular weight of their protein components than do eubacterial 50S particles. 5. Particle weights estimated by either buoyant-density data, or molecular weights of rRNA plus protein, agree to within less than 2%. By either procedure C. acidophila 30S subunits 1.15×10(6) Da mass) are estimated to be about 300 000 Da heavier than their eubacterial counterparts (0.87×10(6) Da mass); a smaller difference. 0.15×10(6) Da, exists between the archaebacterial and the eubacterial 50S subunits (respectively 1.8×10(6) and 1.65×10(6) Da). It is concluded that the heavier-than-eubacterial mass of the C. acidophila ribosomes resides principally in their smaller subunits.

scholarly journals Decoding regulatory specificity of human ribosomal proteins on global gene expression

2021 ◽  
Author(s):  
Yizhao Luan ◽  
Nan Tang ◽  
Jiaqi Yang ◽  
Shuting Liu ◽  
Chichi Cheng ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Ribosomal Proteins ◽  
Translational Regulation ◽  
Large Subunit ◽  
Global Gene Expression ◽  
Small Subunit ◽  
Sequencing Analysis ◽  
Retina Development ◽  
P53 Signaling

Human ribosomes, made of around 80 ribosomal proteins (RPs) and four ribosomal RNAs, have long been thought as uniform passive protein-making factories with few regulatory functions. Recently, accumulating evidence showed heterogeneity of RP composition in ribosomes responsible for regulating gene expression in development and tumorigenesis. However, a comprehensive understanding of regulatory spectrum of RPs is unclear. In this study, we conducted a systematic survey of regulatory specificity of human RPs on global gene expression. We assessed deficiency of 75 RP, including 44 from the large subunit (60S) and 31 from the small subunit (40S), on gene translation and transcription via ribosomal profiling and RNA sequencing analysis. We showed that RP deficiency induced diverse expression changes, particularly at the translational level. RPs were subjected to co-translational regulation under ribosomal stress where deficiency of the 60S or the 40S RPs had distinguished effects on the two subunits. The gene ontology analysis revealed that RP deficiency perturbed expression of genes related to cell cycle, cellular metabolism, signal transduction and development. Deficiency of RPs from the 60S led to a greater repression effect on cell growth than that from the 40S by affecting P53 signaling and cell cycle pathways. To demonstrate functional specificity of RPs, we showed that RPS8 deficiency stimulated cellular apoptosis but RPL13 or RPL18 deficiency promoted cellular senescence. We also showed that RPL11 and RPL15 played important roles in retina development and angiogenesis, respectively. Overall, our study demonstrated a widespread regulatory role of RPs in controlling cellular activity and provided an important resource which offered novel insights into ribosome regulation in human diseases and cancer.

scholarly journals Some Physicochemical Properties of Erysimum Latent Virus

1982 ◽  
Vol 35 (1) ◽  
pp. 5
Author(s):  
Keith H Gough ◽  
Glenn G Lilley ◽  
Dharma D Shukla ◽  
Frank Woods
Keyword(s):  
Molecular Weight ◽  
Buoyant Density ◽  
Latent Virus ◽  
Sedimentation Equilibrium ◽  
Protein Subunit ◽  
Molecular Weights ◽  
Caesium Chloride ◽  
Rna Content ◽  
Velocity Diffusion ◽  
Phosphorus Determination

Sedimentation velocity, diffusion coefficient and sedimentation equilibrium measurements gave a molecular weight of 5 �90 x 106 for the intact Erysimum latent virus. The molecular weight of the empty shell was estimated to be 3�92 X 106 and the protein subunit to be 21 600. The RNA content calculated from the molecular weights of the full and empty particles is 33 %, in agreement with that estimated from the buoyant density in caesium chloride. However, a direct phosphorus determination gave an RNA content of only 28 %.

Creation of a Ribosomal Protein Sa/LAMR1 Heterozygous Mouse.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3551-3551
Author(s):  
Steven R. Ellis ◽  
Paula M. Logsdon ◽  
Junying Han
Keyword(s):  
Ribosomal Protein ◽  
Ribosomal Proteins ◽  
Small Subunit ◽  
Fourth Generation ◽  
Unknown Etiology ◽  
Gene Encoding ◽  
Mendelian Segregation ◽  
Ribosomal Subunits ◽  
Premature Deaths ◽  
Ribosomal Protein S19

Abstract Diamond Blackfan anemia (DBA) is a severe hypoplastic anemia that generally presents early in infancy. Approximately 25% of DBA cases have been linked to mutations in the gene encoding ribosomal protein S19. The remaining cases are of unknown etiology. Our studies in yeast have identified a specific role for Rps19 in the maturation of 40S ribosomal subunits. While only one other small subunit ribosomal protein, Rps18, has a function virtually identical to Rps19 in subunit maturation, several others have functions closely related to Rps19. If the involvement of Rps19 in DBA is linked to its role in the synthesis of 40S ribosomal subunits, we would expect that one or more of these other ribosomal proteins may be responsible for DBA in patients with normal RPS19. To address a potential role for ribosomal proteins other than Rps19 in DBA we have created a transgenic mouse heterozygous at the LAMR1 locus. LAMR1 encodes ribosomal protein Sa, the mammalian homolog of the yeast ribosomal protein S0. Rps0, like Rps19, is required for the maturation of the 3′ end of 18S rRNA. We are currently in the fourth generation of out-crossing the original 129SvEv/C57BL6J chimeras to a C57BL6 background. The frequency of heterozygous pups is about that expected by Mendelian segregation suggesting that haploinsufficiency for ribosomal protein Sa does not lead to significant embryonic lethality. The heterozygous mice do, however, exhibit a higher frequency of craniofacial abnormalities and premature deaths relative to their wild-type littermates. The overall fitness of the heterozygous mice appears to be decreasing with each generation of outcrossing to the C57/BL6 background. Efforts are underway to understand the nature of the premature deaths and to obtain detailed hematological profiles on the LAMR1 heterozygous mice.

scholarly journals Functional dichotomy of ribosomal proteins during the synthesis of mammalian 40S ribosomal subunits

2010 ◽  
Vol 190 (5) ◽  
pp. 853-866 ◽  
Author(s):  
Marie-Françoise O’Donohue ◽  
Valérie Choesmel ◽  
Marlène Faubladier ◽  
Gwennaële Fichant ◽  
Pierre-Emmanuel Gleizes
Keyword(s):  
Nuclear Export ◽  
Ribosomal Proteins ◽  
Small Subunit ◽  
The Body ◽  
Subunit Structure ◽  
Rrna Processing ◽  
Ribosomal Subunits ◽  
Diamond Blackfan Anemia ◽  
Nucleolar Chromatin ◽  
Cytoplasmic Maturation

Our knowledge of the functions of metazoan ribosomal proteins in ribosome synthesis remains fragmentary. Using siRNAs, we show that knockdown of 31 of the 32 ribosomal proteins of the human 40S subunit (ribosomal protein of the small subunit [RPS]) strongly affects pre–ribosomal RNA (rRNA) processing, which often correlates with nucleolar chromatin disorganization. 16 RPSs are strictly required for initiating processing of the sequences flanking the 18S rRNA in the pre-rRNA except at the metazoan-specific early cleavage site. The remaining 16 proteins are necessary for progression of the nuclear and cytoplasmic maturation steps and for nuclear export. Distribution of these two subsets of RPSs in the 40S subunit structure argues for a tight dependence of pre-rRNA processing initiation on the folding of both the body and the head of the forming subunit. Interestingly, the functional dichotomy of RPS proteins reported in this study is correlated with the mutation frequency of RPS genes in Diamond-Blackfan anemia.

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