Eukaryotic protein synthesis

Rhoads, R. E. (1999). Minireview Signal transduction pathways that regulate eukaryotic protein synthesis. J. Biol. Chem. 274, 30337-30340. 

Lamphear, B. J., Kirchweger, R., Skem, T., and Rhoads, R. E. (1995). Mapping of functional domains in eukaryotic protein synthesis initiation factor 4G (eIF4G) with picornaviral proteases. Implications for cap-dependent and cap-independent translational initiation. J. Biol. Chem. 270, 21975—21983. 

Cundliffe, E., Cannon, M., and Davies, J. (1974). Mechanism of inhibition of eukaryotic protein synthesis b trichothecene fungal toxins. Proc. Natl. Acad. Sci. USA 71, 30-34. 

Alice et al studied the turnover kinetics of Listeria OTonocytogenex-secreted p60 protein (a murein hydrolase) by host cell cytosolic proteasomes. J774 cells, seeded in flasks and incubated overnight in culture medium, were infected with log-phase cultures of E. monocytogenes for 30 min, washed, and incubated in culture medium for 3 h, with gentamicin (50 tg/ml) added after the first 30 min to inhibit extracellular bacterial growth. Cells then were washed and placed in methionine-free medium with spectinomycin, gentamicin, the eukaryotic protein synthesis inhibitors [cycloheximide (50 tg/mL) and anisomycin (30 tg/ml),] and 25 dVI calpain inhibitor I. After 30 min, [ S]methionine was added, and the cells were pulse-labeled for periods of 20 to 60 min. Cells... 

The emetines include emetine and dehydroemetine. These drugs act only against trophozoites. Their mechanism of action is based on eukaryote protein synthesis. They are parenterally administered because oral preparations are absorbed erratically and may induce severe vomiting. They are widely distributed and accumulate in liver, lungs and other tissues. The emetines are slowly elimination via the kidneys. Local side-effects in the area of the intramuscular injection are pain, tenderness and muscle weakness. Serious toxicity is common if the drugs are given for more than 10 days. Side effects include nausea, vomiting, diarrhoea but also cardiotoxicity. 

An understanding of protein synthesis, the most complex biosynthetic process, has been one of the greatest challenges in biochemistry. Eukaryotic protein synthesis involves more than 70 different ribosomal proteins 20 or more enzymes to activate the amino acid precursors a dozen or more auxiliary enzymes and other protein factors for the initiation, elongation, and termination of polypeptides perhaps 100 additional enzymes for the final processing of different proteins and 40 or more kinds of transfer and ribosomal RNAs. Overall, almost 300 different macromolecules cooperate to synthesize polypeptides. Many of these macromolecules are organized into the complex three-dimensional structure of the ribosome. 

Protein synthesis is a central function in cellular physiology and is the primary target of many naturally occurring antibiotics and toxins. Except as noted, these antibiotics inhibit protein synthesis in bacteria. The differences between bacterial and eukaryotic protein synthesis, though in some cases subtle, are sufficient that most of the compounds discussed below are relatively harmless to eukaryotic cells. Natural selection has favored the evolution of compounds that exploit minor differences in order to affect bacterial systems selectively, such that these biochemical weapons are synthesized by some microorganisms and are extremely toxic to others. Because nearly every step in protein synthesis can be specifically inhibited by one antibiotic or another, antibiotics have become valuable tools in the study of protein biosynthesis. 

The pathway of protein synthesis translates the three-letter alphabet of nucleotide sequences on mRNA into the twenty-letter alphabet of amino acids that constitute proteins. The mRNA is translated from its 5 -end to its 3 -end, producing a protein synthesized from its amino-terminal end to its carboxyl-terminal end. Prokaryotic mRNAs often have several coding regions, that is, they are polycistronic (see p. 420). Each coding region has its own initiation codon and produces a separate species of polypeptide. In contrast, each eukaryotic mRNA codes for only one polypeptide chain, that is, it is monocistronic. The process of translation is divided into three separate steps initiation, elongation, and termination. The polypeptide chains produced may be modified by posttranslational modification. Eukaryotic protein synthesis resembles that of prokaryotes in most details. [Note Individual differences are mentioned in the text.]... 

Figure 29-11 Initiation of eukaryotic protein synthesis. 1, 2, 3 = elFl S...

The binding of interferons to their receptors induces a rapid increase in the transcription of particular genes and synthesis of corresponding proteins.196 202 One of the proteins induced is a double-stranded RNA-activated 2 -5 A synthase, which polymerizes ATP to a series of 2 -5 linked oligonucleotides containing triphosphates at the 5 termini.202-204 Double-stranded RNA is uncommon except in replicating viruses, and it is thought that the activation by dsRNA is related to establishment of an antiviral state. Another interferon-induced enzyme is the small subunit of eukaryotic protein synthesis initiation factor eIF-2. 

Robert F, Gao HQ, Donia M, Merrick WC, Hamann MT, Pelletier J (2006) Chlorolissocli-mides New Inhibitors of Eukaryotic Protein Synthesis. RNA 12 717... 

The broad outlines of eukaryotic protein synthesis are the same as in prokaryotic protein synthesis. The genetic code is generally the same (some microorganisms and eukaryotic mitochondria use slightly different codons), rRNA and protein sequences are recognizably similar, and the same set of amino acids is used in all organisms. However, specific differences exist between the two types of protein synthesis at all steps of the process. 

Rhoads RE. Regulation of eukaryotic protein synthesis by initiation factors. J Biol Chem 268 3017-3020, 1993. 

High concentrations of hemin inhibit the transport of ALA synthase into the mitochondria, where one of the substrates, succinyl-CoA, is formed. Thus, heme synthesis is inhibited until enough globin is made to react with any heme already formed. Low concentrations, or the absence, of hemin is the signal that globin is not needed this protein (and, therefore, globin) synthesis is inhibited. In the absence of hemin, a protein kinase is activated this phosphorylates an initiation factor of (eukaryotic) protein synthesis, eIF-2, which then inhibits polypeptide chain initiation (Chap. 17) and hence inhibits globin synthesis. 

Ageladine A (132), which is a 9-methyladenine derivative of diterpene isolated from a sponge Agelas sp., is antimicrobial and inhibitory against Na, K-ATPase (12). A novel, chlorinated diterpenoid, chlorolissoclimide (133), which was isolated from the ascidian Lissoclinum forskalii, inhibits eukaryotic protein synthesis (38). 

Eukaryotic Protein Synthesis Differs from Prokaryotic Protein Synthesis Primarily in Translation Initiation... 

The basic plan of protein synthesis in eukaryotes and archaea is similar to that in bacteria. The major structural and mechanistic themes recur in all domains of life. However, eukaryotic protein synthesis entails more protein components than does prokaryotic protein synthesis, and some steps are more intricate. Some noteworthy similarities and differences are as follows ... 

M. Bushell, W. Wood, M.J. Clemens, and S.J. Morley. 2000. Changes in integrity and association of eukaryotic protein synthesis initiation factors during apoptosis Eur. J. Biochem. 267 1083-1091. (PubMed)... 

T.V. Pestova and C.U. Hellen. 2000. The structure and function of initiation factors in eukaryotic protein synthesis Cell Mol. Life Sci. 57 651-674. (PubMed)... 

Wilhelm JM, Jessop JJ, Pettitt SE, Aminoglycoside antibiotics and eukaryotic protein synthesis stimulation of errors in the translation of natural messengers in extracts of cultured human cells, Biochem, 1978,17(7) 1149-53. 

A novel derivative of histidine present only in the eukaryotic protein elongation factor 2 (EF-2), which participates in the elongation step of protein biosynthesis. Diphtheria toxin inhibits eukaryotic protein synthesis by catalyzing a covalent modification of diphthamide (see Chapter 25). 

Shiga toxin produced by Shigella dysenteriae has similar structural features. The toxin binds to a glycolipid (Gb3), undergoes endocytosis, and the enzymatie Ai fragment, which is a specific N-glycosidase, removes adenine from one particular adenosine residue in the 28S RNA of the 60S ribosomal subunit. Removal of the adenine inactivates the 60S ribosome, blocking protein synthesis. Ricin, abrin, and a number of related plant proteins inhibit eukaryotic protein synthesis in a similar manner (Chapter 25). 

The actions of diphtheria and pertussis toxins are also mediated by ADP-ribosylation. Diphtheria toxin inhibits eukaryotic protein synthesis by ADP ribosylation of elongation factor II (Chapter 23). Pertussis toxin inactivates Gi by ADP ribosylation of its A-subunit and causes an increase in cAMP production. Unlike cholera toxin, pertussis and diphtheria toxins gain access to many tissues to produce diverse biological effects. Severe watery diarrhea... 

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