Ribosomal Protein Conferring Sensitivity to the Antibiotic Spectinomycin in Escherichia coli

Reconstitution of 30S ribosomal particles from 16S ribosomal RNA and total proteins, or from core proteins and split proteins obtained from the ribosomes of strains of Escherichia coli sensitive to and resistant to spectinomycin, shows that the split protein fraction determines the response of polypeptide synthesis in virto to spectinomycin. Reconstitution of active particles in the presence of isolated split proteins allowed the identification of the single split protein responsible for spectinomycin sensitivity.     

Altered ribosomal protein in streptomycin-dependent Escherichia coli

We have compared the 30S ribosomal proteins of strains of Escherichia coli sensitive to and dependent on streptomycin and identified a single protein that is functionally altered in the ribosomes dependent on streptomycin. This protein (30S-15) is the same protein that is functionally altered in ribosomes resistant to streptomycin.     

Amino acid changes provoked by streptomycin in a polypeptide synthesized in vitro

The misincorporation of isoleucine provoked by streptomycin in a polyuridylate-directed incorporating system from Escherichia coli has been examined by a double-labeling technique with phenylalanine-(3)H and isoleucine-(14)C. The polypeptides synthesized are associated with 70S ribosomes and contain only phenylalanine in the absence of streptomycin, and both phenylalanine and isoleucine in the presence of the antibiotic. Results of acid hydrolysis and subsequent chymotryptic digestion of the polypeptides indicate that the misincorporated amino acid is inserted by peptide bonding with phenylalanine and is randomly distributed along the chain.     

Phenotype masking and streptomycin dependence

In attempting to define the role of ribosomes in the mechanism of streptomycin dependence, a new phenomenon has been discovered. Analysis of this phenomenon-called phenotypic masking-leads to the conclusion that "streptomycin dependent" mutants are actually "drug dependent" because their dependence is equally satisfied by several drugs. These drugs, some of which are totally unrelated chemically, act on the ribosome and induce misreading in vitro and suppression in vivo.     

散养鸡鼠伤寒沙门氏菌的分离?鉴定?耐药性研究

从疑似感染的成年散养鸡蛋壳和发病雏鸡的肝脏、脾脏中分离到2株细菌,通过培养特性、镜检结果、生化鉴定、血清学分型等,2株分离细菌均被鉴定为鼠伤寒沙门氏菌,分别命名为AH-DY01和AH-DY02。药物敏感性检测结果表明,AH-DY01对氟苯尼考、左氧氟沙星和环丙沙星敏感,对其他12种抗生素耐药;AH-DY02对阿米卡星、氟苯尼考、左氧氟沙星、复方新诺明、庆大霉素、链霉素、环丙沙星和大观霉素敏感,对其他7种药物耐药,2株沙门氏菌均表现多重耐药性。动物回归试验结果表明,2株鼠伤寒沙门氏菌均可致死雏鸡。对分离菌的16S r DNA进行测序与遗传进化分析,结果表明AH-DY01与标准菌株ATCC-1592和ATCC13311的同源性最高,而AH-DY02与日本分离株之间的亲缘关系最近。     

Cytoplasmic and chloroplast ribosomes of Chlamydomonas: ultracentrifugal characterization

Ribosomes isolated from the cytoplasmic and chloroplast fractions of Chlamydomonas were characterized in the ultracentrifuge. The cytoplasmic ribosomes belong to the 80S class of ribosomes, and, like animal ribosomes, dissociate to 60, 50, and 40S subunits. However, like the ribosomes of microorganisms, they contain smaller RNA's, 24 and 16S, and require 0.01 mole of magnesium ions per liter for stability. Chloroplast ribosomes are 70S like those of higher plants but are very unstable. A stable 50S subunit has been observed.     

RIBOSOMAL RNA ON THE SURFACE OF RIBOSOMES

Ribonuclease from pancreas releases nucleotide from Escherichiacoli ribosomes while not altering significantly the base composition of the total ribosomal RNA. Ribonuclease hydrolyzes ribsomal RNA without destroying the structure of 70S or 30S and 50S ribosomes. The RNA in 30S and 50S ribosomes appears more sensitive to the action of ribonuclease. The data suggest that RNA may be a surface component of the ribosome.     

Structure of RNA in ribosomes

The 50S and 30S ribosomes and 23S and 16S RNA were hydrolyzed with ribonuclease A. The rate constants and number of fragments produced were determined for each reaction. The conformation of 23S RNA changes when the RNA is extracted from the ribosome. Specific regions of the RNA in 50S and 30S ribosomes are protected from hydrolysis by the ribosomal proteins.     

Heterogeneity of 5S RNA in fungal ribosomes

Neurospora crassa has at least seven types of 5S RNA genes (alpha, beta, gamma, epsilon, delta, zeta, and eta) with different coding regions. A high resolution gel electrophoresis system was developed to separate minor 5S RNA's from the major 5S RNA (alpha). A study of several Neurospora crassa strains, four other species in the genus Neurospora, members of two closely related genera, and three distantly related genera demonstrated that 5S RNA heterogeneity is common among fungi. In addition, different 5S RNA's are present in Neurospora ribosomes. The finding that fungal ribosomes are structurally heterogeneous suggests that ribosomes may be functionally heterogeneous as well.     

Reticulocyte protein synthesis: response of ribosome fractions to polyuridylic acid

Reticulocyte ribosomes with sedimentation coefficients greater than 100S ("heavy" ribosomes) appear to be considerably more active in hemoglobin synthesis than are 78S ribosomes. When assayed for the ability to synthesize polyphenylalanine in the presence of polyuridylic acid, the 78S ribosomes and "heavy" ribosomes have similar activities. Polyuridylic acid inhibits incorporation by "heavy" ribosomes of amino acids other than phenylalanine.     

DNA Complementary to Ribosomal RNA: Relation between Genomic Proportion and Ploidy

Ten Nicotiana species were assayed for the proportion of DNA that is complementary to ribosomal RNA. This proportion varies from 0.27 to 0.9 percent, with tetraploid species having lower values than the diploid species. The tetraploid species have about twice as much DNA per cell as do diploid species. Thus, the absolute number of genes for ribosomal RNA varies less than the proportion of complementary DNA. Further, the number of genes for the RNA in 80S ribosomes varies less among species than does that for the RNA in 70S ribosomes. The data indicate that loss of DNA complementary to ribosomal RNA is associated with tetraploidy in the genus Nicotiana.     

Mendelian and uniparental alterations in erythromycin binding by plastid ribosomes

Erythromycin binds specifically to the 52S subunit of the chloroplast ribosome of Chlamydomonas reinhardi. A number of erythromycin-resistant mutants whose ribosomes have lost their affinity for the antibiotic have been isolated, but the sedimentation properties of their ribosomes are indistinguishable from those of the wild-type strain. These mutants represent at least three genetic loci. Two of them show Mendelian inheritance, and one of them is inherited in a uniparental manner.     

一例引起史氏鲟鱼创口感染的病原菌分离与鉴定

分离和鉴定引起史氏鲟鱼创口感染的病原菌,为史氏鲟鱼疾病防治提供科学依据。本实验分析了一例刚捕获的患有经微创手术后创口感染疾病的史氏鲟腐败肉和肝脏的细菌菌相,分离获得优势菌株YSJ01,对优势菌进行了形态学、药物敏感性试验、动物感染试验和VITEK全自动微生物分析系统鉴定。结果显示：菌株YSJ01为革兰氏阳性菌;VITEK全自动微生物系统鉴定菌株YSJ01为致病性嗜水气单胞菌;动物感染试验证明：史氏鲟经创伤感染后表现为创口溃烂的典型症状;药敏试验结果表明：菌株YSJ01对氟苯尼考、复方新诺明、大观霉素、利福平、强力霉素、卡那霉素等9种药物敏感。对青霉素、氨苄西林、羧苄青霉素、万古霉素、新生霉素、土霉素、萘啶酸、阿莫西林、林可霉素等表现出耐药性。本研究为南方史氏鲟鱼经微创手术后创口感染疾病的防治提供了重要的实验依据。     

Carbon tetrachloride poisoning in rats: alteration in ribosomes of the liver

Microsomes from livers of albino rats treated with carbon tetrachloride were compared with those from normal rats with respect to their ability to incorporate amino acid. Acute carbon tetrachloride poisoning results in depressed capacity of microsomes to incorporate amino acid. From ultracentrifugal data, there is an apparent dissociation of 79S ribosomes into 54S components.     

Erythromycin-resistant mutant of Escherichia coli with altered ribosomal protein component

Erythromycin combines with 50S ribosomal subunit of an erythromycin-sensitive Escherichia coli (strain Q13), while ribosomes from an erythromycin-resistant mutant from this strain have little affinity for the antibiotic. A protein component of the 50S subunit of the mutant strain is distinct from that of the parent Q13 strain.     

Streptomycin resistance mutation in Escherichia coli: altered ribosomal protein

Reconstitution of 30S ribosomal particles was performed with 16S ribosomal RNA, "core" proteins, and "split" proteins from 30S particles derived from streptomycin-sensitive and streptomycin-resistant Escherichia coli cells in various combinations. Analysis of streptomycin sensitivity of the reconstituted particles has shown that the alteration induced by the resistance mutation resides in the core proteins, and not in the RNA or in the split proteins of the 30S particles.     

Functional chloroplast polyribosomes from tobacco leaves

Incubation of isolated to bacco chloroplasts with ingredients re quired for protein synthesis resulted in liberation of 70S ribosomnes and poly ribosomes that no longer sedimented with the chloroplasts. With increasing time of incubation, polyribosomes broke down to 70S monosomes. Similarly, microgram quantities of ribonuclease caused chloroplast polyribosomes to break down into monosomes. Both polyribosomes and 70S ribosomes that were isolated on sucrose density gra dients and tested separately in cell-free systems were capable of protein syn thesis; however, polyribosomes formed more protein per unit of RNA than monosomes did.     

兔巴氏杆菌与大肠杆菌混合感染病例诊断及药敏试验

为了诊断患病死亡兔,对死亡兔进行临床解剖,细菌分离培养,获得DX01、DX02、JX01、JX02等4株菌.按照常规生化鉴定法,对分离菌进行生理生化实验,结果:DX01、DX02对甘露醇、乳糖、葡萄糖、麦芽糖均产酸产气,不发酵蔗糖,枸橼酸钠、尿素、V-P、硫化氢、明胶等反应均为阴性,靛基质、M-R、硝酸盐等反应均为阳性,与大肠杆菌标准株完全一致;JX01、JX02均发酵甘露醇、蔗糖、葡萄糖、麦芽糖,不发酵乳糖,靛基质、M-R、V-P、硫化氢、明胶等反应均为阴性,硝酸盐反应为阳性,与巴氏杆菌标准株基本一致,JX01、JX02进一步经过16SrDNA检测与测序分析,与巴氏杆菌同源性为100％.分离菌动物接种试验结果,试验组小白鼠在2日内均死亡.死亡小白鼠体内均分离出了原培养物.结论:DX01、DX02为大肠杆菌,JX01、JX02为大肠杆菌,患病死亡兔由大肠杆菌和巴氏杆菌混合感染引起的.按照琼脂扩散法做药敏实验,结果DX01、DX02只对卡那霉素、氧氟沙星、甲氧嘧啶敏感,对其他的药物不敏感;JX01、JX02对氧氟沙星、甲氧嘧啶、庆大霉素、呋喃唑酮、强力霉素、链霉素、复方新诺明、大观霉素等敏感,对头孢拉啶等中度敏感.治疗该病临床用药首选氧氟沙星、甲氧嘧啶.     

Crystal structure of the eukaryotic 60S ribosomal subunit in complex with initiation factor 6

Protein synthesis in all organisms is catalyzed by ribosomes. In comparison to their prokaryotic counterparts, eukaryotic ribosomes are considerably larger and are subject to more complex regulation. The large ribosomal subunit (60S) catalyzes peptide bond formation and contains the nascent polypeptide exit tunnel. We present the structure of the 60S ribosomal subunit from Tetrahymena thermophila in complex with eukaryotic initiation factor 6 (eIF6), cocrystallized with the antibiotic cycloheximide (a eukaryotic-specific inhibitor of protein synthesis), at a resolution of 3.5 angstroms. The structure illustrates the complex functional architecture of the eukaryotic 60S subunit, which comprises an intricate network of interactions between eukaryotic-specific ribosomal protein features and RNA expansion segments. It reveals the roles of eukaryotic ribosomal protein elements in the stabilization of the active site and the extent of eukaryotic-specific differences in other functional regions of the subunit. Furthermore, it elucidates the molecular basis of the interaction with eIF6 and provides a structural framework for further studies of ribosome-associated diseases and the role of the 60S subunit in the initiation of protein synthesis.     

Structure of Ribosomal Subunits of M. vannielii: Ribosomal Morphology as a Phylogenetic Marker

On the basis of ribosomal morphology, it has been proposed that the sulfur-metabolizing archaebacteria constitute a group (the eocytes) with a phylogenetic importance equal to that of the eubacteria, archaebacteria, and eukaryotes. It has been further proposed that eocytes should be given kingdom status. Ribosomal subunits from the methanogenic archaebacterium Methanococcus vannielii were examined by electron microscopy, and their structures were compared to those of other archaebacterial, eubacterial, and eukaryotic ribosomes. 30S subunits from M. vannielii showed the elongated contour and the one-third to two-thirds partition characteristic of such subunits. In addition, the angled asymmetric projections of those subunits showed a squarish base and a beak on the head. 50S subunits from M. vannielii were seen in both crown and kidney views. In crown views, the L1 protuberance was frequently pronounced and split; an incision below this protuberance and a protrusion at the base of the particle were also observed. Although previous studies suggested that certain of these structural features were found exclusively in ribosomes from sulfur-metabolizing archaebacteria, these new results indicate that such features also occur in ribosomes from a typical methanogenic archaebacterium and thus may not be reliable phylogenetic markers.