Site-specific cleavage of the 40S ribosomal subunit reveals eukaryote-specific ribosomal protein S28 in the subunit head

After resolving the crystal structure of the prokaryotic ribosome, mapping the proteins in the eukaryotic ribosome is a challenging task. We applied RNase H digestion to split the human 40S ribosomal subunit into head and body parts. Mass spectrometry of the proteins in the 40S subunit head revealed the presence of eukaryote-specific ribosomal protein S28e. Recombinant S28e was capable of specific binding to the 3′ major domain of the 18S rRNA (K_a = 8.0 ± 0.5 × 10⁹ M⁻¹). We conclude that S28e has a binding site on the 18S rRNA within the 40S subunit head.

Structured summary

MINT-8044084: S8 (uniprotkb:P62241) and S19 (uniprotkb:P39019) colocalize (MI:0403) by cosedimentation through density gradient (MI:0029)MINT-8044095: S8 (uniprotkb:P62241), S19 (uniprotkb:P39019) and S13 (uniprotkb:P62277) colocalize (MI:0403) by cosedimentation through density gradient (MI:0029)MINT-8044024: S29 (uniprotkb:P62273), S28 (uniprotkb:P62857), S21 (uniprotkb:P63220), S20 (uniprotkb:P60866), S26 (uniprotkb:P62854), S25 (uniprotkb:P62851), S12 (uniprotkb:P25398), S17 (uniprotkb:P08708), S19 (uniprotkb:P39019), S14 (uniprotkb:P62263), S16 (uniprotkb:P62249) and S11 (uniprotkb:P62280) colocalize (MI:0403) by cosedimentation through density gradient (MI:0029)MINT-8044065: S29 (uniprotkb:P62273), S28 (uniprotkb:P62857), S19 (uniprotkb:P39019), S14 (uniprotkb:P62263) and S16 (uniprotkb:P62249) colocalize (MI:0403) by cosedimentation through density gradient (MI:0029)     

Twelve newSalmonella types

Twelve hitherto unrecognizedSalmonella types are described. S. heerlen (11 : i : 1,6) andS. sloterdijk (1, 4, 12, 27 : z 35 : z 6) were isolated from patients in the Netherlands,S. maartensdijk (40 : g, p) was isolated from a healthy calf,S. maastricht (11 : z 41 : 1, 2(7)) from imported fishmeal,S. parera (11 : z 4, z 23 : -.) from water on the island Bonaire,S. putten (13, 23, 36 : d : 1, w),S. hoograven (50 : z 10 : z 6 : z 42),S. schalkwijk ((6), 14, (24) : i : e, n, ...),S. hilversum (30 : k : 1, 2) andS. harmelen (51 : z 4, z 23 : -.) from reptiles,S. breukelen (6, 8 : 1, z 13 : e, n, z 15) from a cuscus andS. maarssen (9, 46 : z 4, z 24 : z 39 : z 42) from a lizard.     

Modulation of Platelet Activation and Thrombus Formation Using a Pan-PI3K Inhibitor S14161

The phosphatidylinositol 3–kinase (PI3K) signaling pathway is critical in modulating platelet functions. In the present study, we evaluated the effect of {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161, a recently identified pan-class I PI3K inhibitor, on platelet activation and thrombus formation. Results showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 inhibited human platelet aggregation induced by collagen, thrombin, U46619, and ADP in a dose-dependent manner. Flow cytometric studies showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 inhibited convulxin- or thrombin-induced P-selectin expression and fibrinogen binding of single platelet. {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 also inhibited platelet spreading on fibrinogen and clot retraction, processes mediated by outside-in signaling. Using a microfluidic chamber we demonstrated that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 decreased platelet adhesion on collagen-coated surface by about 80%. Western blot showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 inhibited phosphorylation of Akt at both Ser473 and Thr308 sites, and GSK3β at Ser9 in response to collagen, thrombin, or U46619. Comparable studies showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 has a higher potential bioavailability than {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}}LY294002, a prototypical inhibitor of pan-class I PI3K. Finally, the effects of {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 on thrombus formation in vivo were measured using a ferric chloride-induced carotid artery injury model in mice. The intraperitoneal injection of {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 (2 mg/kg) to male C57BL/6 mice significantly extended the first occlusion time (5.05±0.99 min, n = 9) compared to the vehicle controls (3.72±0.95 min, n = 8) (P<0.05), but did not prolong the bleeding time (P>0.05). Taken together, our data showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 inhibits platelet activation and thrombus formation without significant bleeding tendency and toxicity, and considering its potential higher bioavailability, it may be developed as a novel therapeutic agent for the prevention of thrombotic disorders.     

BRCA2 is needed for both repair and cell cycle arrest in mammalian cells exposed to S23906, an anticancer monofunctional DNA binder

Repair of DNA-targeted anticancer agents is an active area of investigation of both fundamental and clinical interest. However, most studies have focused on a small number of compounds limiting our understanding of both DNA repair and the DNA damage response. {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 is an acronycine derivative that shows strong activity toward solid tumors in experimental models. {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 forms bulky monofunctional DNA adducts in the minor groove which leads to destabilization of the double-stranded helix. We now report that {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 induces formation of DNA double strand breaks that are processed through homologous recombination (HR) but not Non-Homologous End-Joining (NHEJ) repair. Interestingly, {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 exposure was accompanied by a higher sensitivity of BRCA2-deficient cells compared to other HR deficient cell lines and by an S-phase accumulation in wild-type (wt), but not in BRCA2-deficient cells. Recently, we have shown that {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906-induced S phase arrest was mediated by the checkpoint kinase Chk1. However, its activated phosphorylated form is equally induced by {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 in wt and BRCA2-deficient cells, likely indicating a role for BRCA2 downstream of Chk1. Accordingly, override of the S phase arrest by either 7-hydroxystaurosporine (UCN-01) or AZD7762 potentiates the cytotoxic activity of {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 in wt, but not in BRCA2-deficient cells. Together, our findings suggest that the pronounced sensitivity of BRCA2-deficient cells to {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 is due to both a defective S-phase arrest and the absence of HR repair. Tumors with deficiencies for proteins involved in HR, and BRCA2 in particular, may thus show increased sensitivity to {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906, thereby providing a rationale for patient selection in clinical trials.     

NeueSilene-Arten (Caryophyllaceae) aus dem Gebiet der Flora Iranica

15 new species are described: Sect.Sclerocalycinae:S. farsistanica, S. stapfii. — Sect.Spergulifoliae:S. paktiensis. — Sect.Auriculatae:S. caroli-henrici, S. daënensis, S. gertraudiae, S. nizvana, S. oligophylla, S. persepolitana, S. pseudaucheriana, S. pseudonurensis, S. renzii, S. salangensis, S. sojakii. — Sect.Brachypodae:S. rasvandica. — All the new species are from Iran with exception ofS. paktiensis andS. parvanica which are from Afghanistan.

Florae Iranicae praecursores 46–60. — Praecursores praecurrentes in Pl. Syst. Evol.142: 239–246 (1983).     

Six newSalmonella types,isolated in ghana (S. volta,S. agona,S. wa,S. techimani,S. mampong andS. tafo)

Summary Six newSalmonella types isolated in Ghana are described.S. volta, 11: 4: 1,z₁₃, z₂₈ was isolated from a swine;S.agona 4,12: fgs:—,S.wa, 16: b: 1,5S.technimani, 28: c: z₆ andS.tafo, 1, 4 12, 27: z₃₅: 1,7 were isolated from cattle;S.mampong, 13,22: z₃₅: 1,6, was isolated from a lizzard.     

Enterotoxigenicity and invasiveness of Salmonella species

A large number of enterotoxigenic strains was encountered in a group 56 Salmonella cultures belonging to 8 species viz., S. alachua, S. anatum, S. dublin, S. enteritidis, S. hindmarsh, S. newport, S. typhimurium, S. weltevreden, and 5 serotypes of S. arizona (16:z₄:-; 48:1,v:z₅₆; 53:z₅₂:z₅₃; 60:r:z; 61:i:z₅₃). These cultures were isolated mainly from humans and animals suffering from gasteroenteritis. The enterotoxigenic (diarrhoeagenic) Salmonella cultures possess capacities for both skin permeation and epithelial penetration (invasiveness). Preliminary characterization revealed that Salmonella enterotoxin is a heatlabile protein of high molecular weight. It is suggested that enterotoxigenic and invasive propeties play a vital role in the pathogenesis of Salmonella diarrhoea.     

Eight newSalmonella types

Summary Eight newSalmonella types are described.S. houten, 43; Z₄, Z₂₃ :−, was isolated from a frog,S.tuindorp, 43: Z₄, Z₃₂: −,S. vleuten, 44: fg: −,S.zeist, 18: Z₁₀: Z₆, andS.soesterberg. 21: Z₄, Z₂₃: −, were isolated from lizards. S. kralingen (8), 20: y: Z₆, was isolated from peanuts andS. assen, 21: a: −, was isolated from a swine. All these strains were isolated in the Netherlands. S.mim, 13, 22: a: 1,6, was isolated from a rat in Ghana.     

Biochemical methane potential (BMP) test for thickened sludge using anaerobic granular sludge at different inoculum/substrate ratios

The biochemical methane potential (BMP) test for thickened sludge was evaluated at three different inoculum/substrate (I:S) ratios. The cumulative methane yield was 51.4 mL CH₄/g VS_added at an I:S ratio of 1:1, 76.3 mL CH₄/g VS_added at an I:S ratio of 1:3, and 21.9 mL CH₄/g VS_added at an I:S ratio of 1:8. The greatest ultimate methane yield and methane production rate constant were achieved at an I:S ratio of 1:3, whereas the least was obtained at an I:S ratio of 1:8. The maximum methane production rate constant was 0.38/day and the minimum methane production rate constant was 0.0016/day. For the case of a lower I:S ratio, the biomass activity may be affected due to the low substrate concentration. On the other hand, for the case of higher I:S ratios, anaerobic digestion of thickened sludge was inhibited by higher concentrations of volatile fatty acids and lower pH.     

Different effects on triacylglycerol packaging to oil bodies in transgenic rice seeds by specifically eliminating one of their two oleosin isoforms

Expression of OLE16 and OLE18, two oleosin isoforms in oil bodies of rice seeds, was suppressed by RNA interference. Electron microscopy revealed a few large, irregular oil clusters in 35S::ole16i transgenic seed cells, whereas accumulated oil bodies in 35S::ole18i transgenic seed cells were comparable to or slightly larger than those in wild-type seed cells. Large and irregular oil clusters were observed in cells of double mutant seeds. These unexpected differences observed in oil bodies of 35S::ole16i and 35S::ole18i transgenic seeds were further analyzed. In comparison to wild-type plants, OLE18 levels were reduced to approximately 40% when OLE16 was completely eliminated in 35S::ole16i transgenic plants. In contrast, OLE16 was reduced to only 80% of wild-type levels when OLE18 was completely eliminated in 35S::ole18i transgenic plants. While the triacylglycerol content of crude seed extracts of 35S::ole16i and 35S::ole18i transgenic seeds was reduced to approximately 60% and 80%, respectively, triacylglycerol in isolated oil bodies was respectively reduced to 45% and 80% in accordance with the reduction of their oleosin contents. Oil bodies isolated from both 35S::ole16i and 35S::ole18i transgenic seeds were found to be of comparable size and stability to those isolated from wild-type rice seeds, although they were merely sheltered by a single oleosin isoform. The drastic difference between the triacylglycerol contents of crude seed extracts and isolated oil bodies from 35S::ole16i transgenic plants could be attributed to the presence of large, unstable oil clusters that were sheltered by insufficient amounts of oleosin and therefore could not be isolated together with stable oil bodies.     

Stoichiometry of proton release from the catalytic center in photosynthetic water oxidation. Reexamination by a glass electrode study at ph 5.5-7.2.

The catalytic center (CC) of water oxidation in photosystem II passes through four stepwise increased oxidized states (S(0)-S(4)) before O(2) evolution takes place from 2H(2)O in the S(4) --> S(0) transition. The pattern of the release of the four protons from the CC cannot be followed directly in the medium, because proton release from unknown amino acid residues also takes place. However, pH-independent net charge oscillations of 0:0:1:1 in S(0):S(1):S(2):S(3) have been considered as an intrinsic indicator for the H(+) release from the CC. The net charges have been proposed to be created as the charge difference between electron abstraction and H(+) release from the CC. Then the H(+) release from the CC is 1:0:1:2 for the S(0) --> S(1) --> S(2) --> S(3) --> S(0) transition. Strong support for this conclusion is given in this work with the analysis of the pH-dependent pattern of H(+) release in the medium measured directly by a glass electrode between pH 5.5 and 7.2. Improved and crystallizable photosystem II core complexes from the cyanobacterium Synechococcus elongatus were used as material. The pattern can be explained by protons released from the CC with a stoichiometry of 1:0:1:2 and protons from an amino acid group (pK approximately 5.7) that is deprotonated and reprotonated through electrostatic interaction with the oscillating net charges 0:0:1:1 in S(0):S(1):S(2):S(3). Possible water derivatives that circulate through the S states have been named.     

Ribosomal proteins: XLIII. In vivo assembly of Escherichia coli ribosomal proteins

Isolation of ribosomal precursors from Escherichia coli K12 is described. The RNA and protein content of the precursor particles was determined.One physiologically stable precursor was found for the 30 S subunit. The assembly scheme is as follows: p16 S RNA + 9 proteins → p30 S (“21 S” precursor) p30 S + 12 proteins → 30 S subunit where p is precursor.Each of the two precursors for the 50 S subunit, P₁50 S and p₂50 S (“32 S” and “43 S” precursors, respectively), contains p5 S + p23 S RNA's in a 1:1 molar ratio. The assembly scheme is as follows: p23 S RNA + p5 S RNA + 16 or 17 proteins → p₁50 S

In contrast to the p₂50 S precursor the p₁50 S precursor is not similar to any core particles, which were obtained by treating 50 S subunits with different concentrations of LiCl or CsCl.The precursors p30 S and p₂50 S can be converted into active 30 S and 50 S sub-units, respectively, by incubation at 42 °C in the presence of ribosomal proteins and under RNA methylating conditions.     

Dünnschichtchromatographische Bestimmung der Rückstände des Herbicids Leuna MECK im Boden

Abstract

Wetzel, TH. (Edit.): Symposium mit internationaler Beteiligung zur Schaderregerüberwachung in der industriemäßigen Getreideproduktion. 510 S., Teil I bis III, 134 Abb. Halle: 1975. Reviewed by G. Lutze.

O. V.: Pflanzenschutzmittel-Verzeichnis 1974/1975. 1. Aufl. 121 S. Berlin: VEB Dt. Landwirtschaftsverlag 1975. Reviewed by W. Tarnow.

Schumakov, E. M.; Gussev, G. W.; Fedorintschik, N. S. (Edit.): Biologi?eskie sredstva za??ita rastenij. 25000 Aufl. 408 S., 1. Band, 24 Abb. Moskau: Kolos 1974. Reviewed by Th. Wetzel.

Clifton, C. E. (Edit.): Annual Review of Microbiology. 771 S., 25. Band, zahlreiche Abb. Palo Alto: Annual Reviews Inc. 1971. Reviewed by K. Naumann.

Voronkevi?, I. V.: Vy?ivaemost' fitopatogennych Bakterij v Prirode. 268 S., 16 Abb., 41 Tab., Moskau 1974: Nauka. Reviewed by W. Ficke.

Bel'tjukova, K. I.; Koroleva, I. B.; Muras, V. A.: Bakterial'nye bolezni zernobobovych kultur. 339 S., 57 Abb., 49 Tab. Kiev 1974: Naukova Dumka. Reviewed by W. Ficke.

Pospelov, S. M.; Arsen'eva, M. V.; Grusdev, G. S.: Za??ita rastenij. 487 S. Leningrad 1973: Kolos. Reviewed by W. Ficke.

Noordam, D.: Identification of plant viruses. Methods and experiments. 1. Auflage, 218 S., 134 Abb. Wageningen 1973: Centre for agricultural publishing and documentation. Reviewed by D. Spaar.     

Small mammals and their ectoparasites from the Scottish islands of Handa (Sutherland), Muck, Pabay, Scalpay and Soay (Inner Hebrides)

Small mammals were trapped on five islands for short periods during the summers of 1964 and 1965, with the following results:
Handa: Rattus norvegicus only, probably no other species present.
Muck: Sorex araneus, S. minutus, Apodemus sylvaticus and Microtus agrestis; R. norvegicus also present.
Pabay: S. minutus and Neomys fodiens ; probably no other species present.
Scalpay: S. araneus, S. minutus and M. agrestis.
Soay: S. araneus and S. minutus ; rodents almost certainly absent.
Ectoparasites (fleas and Acarina) collected from these small mammals are tabulated.     

Four newSalmonella types (S. gambaga,S. kumasi,S. pramiso andS. ashanti) isolated in Ghana

Summary Four newSalmonella types isolated in Ghana are described.S.gambaga, 21; z35: enz15,S.kumasi, 30;z10: enz15, andS.pramiso, 3,10;c: 1,7 were isolated from reptiles,S. ashanti, 28; b: 1,6 was isolated from a rat.     

Sulfur in Ghanaian soils

Studies were carried out to examine factors which might influence the distribution of S in Ghanaian soils. Nine soil profiles developed over granitic rocks, three each representing the upper slope (US), middle slope (MS) and lower slope (LS) of catena in the evergreen high rain forest (ERF), semi-deciduous rain forest (SDF) and the interior savanna (ISAV) zone of Ghana were selected. The total S contents varied from 9 to 347 ppm; the average for all the surface and subsurface horizons was 141 ppm and for subsoils 105 ppm. The contents also varied according to: (1) the ecological zone as follows: ERF 0) SDF>ISAV and (2) the topographic position: US>MS>LS. The total S was closely correlated with organic C and total N in the surface and subsurface horizons (r=0.931^*** and 0.941^*** respectively). Inorganic sulfate was generally higher in the subsoils than in the surface and subsurface horizons of the ERF and SDF profiles whereas the opposite was the case in the ISAV profiles. Based on the critical value of 6 ppm in surface soils, all the savanna soils would be considered S deficient.The total organic S, which constituted from 56 to over 95% of the total S in the profiles, was significantly correlated with total N both in the surface and subsurface horizons (N:S ratio=9.1:1) and in the subsoils (N:S ratio=7.6:1). Fractionation of the organic S showed that HI-reducible S ranged from 14 to 117 ppm in the surface and subsurface horizons (average 55 ppm, equivalent to 47% of the total organic S) and from 2 to 169 ppm (average 55 ppm, equivalent to 60% of the total organic S) in the subsoils. The C-bonded S ranged from 6 to 223 ppm (average 73 ppm, equivalent to 57% of the total organic S) in the surface and subsurface horizons and from 1 to 83 ppm (average 29 ppm, equivalent to 32% of the total organic S) in the subsoils. HI-reducible S was significantly correlated with organic C (r=0.805^***) and total N (r=0.845^***) in the surface and subsurface horizons only whereas C-bonded S was significantly correlated with organic C and total N in both the surface and subsurface horizons and subsoils (r=0.870^*** and 0.624^*** respectively).The N:S ratios varied from 6.0 to 12.7 in the surface and subsurface horizons and from 0.5 to 27.3 in the subsoils. However the N:S ratio was less variable within the profile than the C:S ratio. The C:N:S ratios varied considerably within the profile and among the different soils but they fall within the range of values reported world-wide.     

Seven newSalmonella types

Seven hitherto unrecognizedSalmonella types are described.S.leiden (13, 22, 36 : z 38),S. enschede (35 : z 10 : l,w) andS. hillegersberg (9, 46 : z 35 : 1,5) were isolated from patients in the Netherlands.S. lawra (44 : k : e,n,z 15) was isolated from a healthy student in Ghana.S. overvecht (30 : a : 1,2),S. woerden (17 : c : z 39) andS. odijk (30 : a : z 39) were isolated from animals kept in zoological gardens in the Netherlands.     

A Comparison of the Native and Derived 30S and 50S Ribosomes of Escherichia coli

Four types of ribosomes occurring in E. coli have been separated by sucrose gradient centrifugation. These are the 30S and 50S particles occurring in E. coli extracts (native particles), and the 30S and 50S particles which are the subunits of 70S ribosomes (derived particles). Two criteria were used in comparing these particles: (1) The type of RNA contained in each, as determined by sedimentation velocity in the analytical ultracentrifuge. (2) The ability of mixtures of 30S and 50S ribosomes (derived 30S + derived 50S, native 30S + native 50S) to undergo the reaction: [Formula: see text] Native and derived 30S particles were found to contain 16S RNA. Derived 50S particles contained 23S RNA and a small amount of 15 to 20S RNA, whereas native 50S ribosomes contained only 16S RNA. Derived 30S and 50S particles combined to form 70S particles. However, under identical conditions, native 30S and 50S particles did not form 70S ribosomes.     

Specific interaction of proteins with 5 S RNA and tRNA in the 42 S storage particle of Xenopus oocytes

During early oogenesis in amphibia, most of the 5 S RNA and tRNA is stored in a ribonucleoprotein particle that sediments at 42 S. In Xenopus laevis the 42 S particle contains two major proteins: of M_r 48 000 (P48) and 43 000 (P43). It is shown that heterogeneity in composition of the 42 S particle reflects a changing situation whereby initially, both 5 S RNA and tRNA are complexed with P48 (1 molecule 5 S RNA: 1 molecule P48; 2 or 3 molecules tRNA: 1 molecule P48), but later, tRNA becomes increasingly associated with P43 (in a 1:1 ratio) although 5 S RNA remains complexed with a cleavage product of P48. These changes relate to the eventual utilization of the excess 5 S RNA and tRNA in ribosome assembly and protein synthesis.