Messenger RNA in early sea-urchin embryos: cytoplasmic particles

Three structures containing messenger RNA can be demonstrated in the cytoplasm of early sea-urchin embryos: (i) particles that sediment more slowly than ribosomes and contain newly synthesized DNA-like RNA, (ii) light polyribosomes, which also contain this newly synthesized RNA, and (iii) heavy polyribosomes, which seemingly contain only already existing or "maternal" messenger RNA and account for the bulk of the synthesis of protein.     

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.     

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.     

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.     

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.     

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.     

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.     

Nucleotide sequence of KB cell 5S RNA

The nucleotide sequence of 5S RNA derived from KB carcinoma cell ribosomes has been determined. The molecule has a length of either 120 or 121 nucleotides with uridine at its 3'-terminus and guanylic acid at its 5'-terminus. If, in addition to Watson-Crick base-pairing, one accepts occasional base-pairing of guanylic acid to uridylic acid, long sequences of complementary nucleotides can be identified within the molecule. Two regions of the molecule contain sequences complementary to four or five bases in the pentanucleotide sequence guanylic acid, ribothymidylic acid, pseudouridylic acid, cytidylic acid, guanylic acid, which is common to most transfer RNA molecules. This is the first time the sequence of an animal-cell RNA has been determined.     

Aurintricarboxylic acid and initiation factors of wheat embryo

In extracts of wheat embryo, aurintricarboxylic acid inhibits attachment of ribosomes to messenger RNA and has little effect on aminoacyl transfer to peptide. This specificity of inhibition allows (i) the demonstration that the initial phase of amino acid incorporation dependent on RNA of tobacco mosaic virus involves an "initiation" reaction between ribosome and messenger RNA which requires two soluble factors, (ii) an assay facilitating purification of the initiation factors, and (iii) the demonstration that these factors are distinct from aminoacyl transfer enzymes.     

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.     

Inhibition of protein synthesis by spectinomycin

Spectinomycin selectively inhibits protein synthesis in cells and in extracts of Escherichia coli. Mutations to high-level resistance to this antibiotic map close to the streptomycin locus, and the site of action of spectinomycin, like that of streptomycin, is the 30S ribosomal subunit, as shown by experiments with reconstituted 70S ribosomes containing subunits from sensitive and from resistant ribosomes. In contrast to streptomycin, however, spectinomycin is not bactericidal and causes no detectable misreading of polyribonucleotides.     

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.     

生草覆盖对红巴梨采后果实主要营养成分变化及重金属元素含量的影响

研究了梨园行间生草覆盖处理对红巴梨果实采后主要营养成分及重金属含量的影响。结果表明,在生草覆盖栽培条件下,红巴梨果实在日平均气温为15～23℃,日平均相对湿度42%～76%的室温环境下贮藏,最佳食用期在第9天。红巴梨采后果实的主要营养成分变化:可溶性固形物的含量在第9天达到最高,峰值为14.27%;还原糖在贮藏的15 d内呈"S"曲线变化,最低值出现在第6天,为7.59%,最高值出现在第12天,为9.22%,随后急剧下降;可溶性总糖从采收后一直处于下降趋势;Vc含量呈双"S"曲线变化,具有先升后降再升再降的变化趋势;粗蛋白含量呈"S"曲线变化,具有先升后降再升的变化趋势,贮藏第9天达到第1个高峰,其含量为258.75 mg/100 g;果实中的7个重金属元素含量均低于绿色鲜梨果(NY/T 423—2000)的要求标准,对重金属元素Cu和Zn的吸收能力强于红香酥和雪花梨。     

Structural basis for the rescue of stalled ribosomes: structure of YaeJ bound to the ribosome

In bacteria, the hybrid transfer-messenger RNA (tmRNA) rescues ribosomes stalled on defective messenger RNAs (mRNAs). However, certain gram-negative bacteria have evolved proteins that are capable of rescuing stalled ribosomes in a tmRNA-independent manner. Here, we report a 3.2 angstrom-resolution crystal structure of the rescue factor YaeJ bound to the Thermus thermophilus 70S ribosome in complex with the initiator tRNA(i)(fMet) and a short mRNA. The structure reveals that the C-terminal tail of YaeJ functions as a sensor to discriminate between stalled and actively translating ribosomes by binding in the mRNA entry channel downstream of the A site between the head and shoulder of the 30S subunit. This allows the N-terminal globular domain to sample different conformations, so that its conserved GGQ motif is optimally positioned to catalyze the hydrolysis of peptidyl-tRNA. This structure gives insights into the mechanism of YaeJ function and provides a basis for understanding how it rescues stalled ribosomes.     

Pyrazinamide inhibits trans-translation in Mycobacterium tuberculosis

Pyrazinamide (PZA) is a first-line tuberculosis drug that plays a unique role in shortening the duration of tuberculosis chemotherapy. PZA is hydrolyzed intracellularly to pyrazinoic acid (POA) by pyrazinamidase (PZase, encoded by pncA), an enzyme frequently lost in PZA-resistant strains, but the target of POA in Mycobacterium tuberculosis has remained elusive. Here, we identify a previously unknown target of POA as the ribosomal protein S1 (RpsA), a vital protein involved in protein translation and the ribosome-sparing process of trans-translation. Three PZA-resistant clinical isolates without pncA mutation harbored RpsA mutations. RpsA overexpression conferred increased PZA resistance, and we confirmed that POA bound to RpsA (but not a clinically identified ΔAla mutant) and subsequently inhibited trans-translation rather than canonical translation. Trans-translation is essential for freeing scarce ribosomes in nonreplicating organisms, and its inhibition may explain the ability of PZA to eradicate persisting organisms.     

How hibernation factors RMF, HPF, and YfiA turn off protein synthesis

Eubacteria inactivate their ribosomes as 100S dimers or 70S monomers upon entry into stationary phase. In Escherichia coli, 100S dimer formation is mediated by ribosome modulation factor (RMF) and hibernation promoting factor (HPF), or alternatively, the YfiA protein inactivates ribosomes as 70S monomers. Here, we present high-resolution crystal structures of the Thermus thermophilus 70S ribosome in complex with each of these stationary-phase factors. The binding site of RMF overlaps with that of the messenger RNA (mRNA) Shine-Dalgarno sequence, which prevents the interaction between the mRNA and the 16S ribosomal RNA. The nearly identical binding sites of HPF and YfiA overlap with those of the mRNA, transfer RNA, and initiation factors, which prevents translation initiation. The binding of RMF and HPF, but not YfiA, to the ribosome induces a conformational change of the 30S head domain that promotes 100S dimer formation.     

青岛市场养殖贝类体内重金属含量的分析

[目的]了解目前青岛市场上贝类的重金属水平。[方法]对主要养殖品种进行随机取样,共采集78个样品。利用湿法消解-石墨炉原子吸收法和等离子体发射光谱法测定重金属Pb、Cd、Cu、Fe、Mn、Zn含量,并采用国家标准对检验结果进行评价。[结果]青岛市场销售的贝类体内6种重金属含量总体较低,仅有部分毛蚶和牡蛎产品中镉含量高于国家标准;养殖贝类的重金属品种间差异较明显。[结论]大部分贝类产品可以安全食用。     

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.     

Crystal structure of the eukaryotic ribosome

Crystal structures of prokaryotic ribosomes have described in detail the universally conserved core of the translation mechanism. However, many facets of the translation process in eukaryotes are not shared with prokaryotes. The crystal structure of the yeast 80S ribosome determined at 4.15 angstrom resolution reveals the higher complexity of eukaryotic ribosomes, which are 40% larger than their bacterial counterparts. Our model shows how eukaryote-specific elements considerably expand the network of interactions within the ribosome and provides insights into eukaryote-specific features of protein synthesis. Our crystals capture the ribosome in the ratcheted state, which is essential for translocation of mRNA and transfer RNA (tRNA), and in which the small ribosomal subunit has rotated with respect to the large subunit. We describe the conformational changes in both ribosomal subunits that are involved in ratcheting and their implications in coordination between the two associated subunits and in mRNA and tRNA translocation.