Role of polyamines in the binding of initiator tRNA to the 70S ribosomes of extreme thermophilic bacterium Calderobacterium hydrogenophilum

1994 ◽  
Vol 161 (6) ◽  
pp. 508-513 ◽  
Author(s):  
Karel Mikulík ◽  
Miroslava Anděrová
Keyword(s):  
Thermophilic Bacterium ◽  
Initiator Trna ◽  
Calderobacterium Hydrogenophilum

scholarly journals The role of histidine-118 of inorganic pyrophosphatase from thermophilic bacterium PS-3

1991 ◽  
Vol 278 (2) ◽  
pp. 595-599 ◽  
Author(s):  
N Hirano ◽  
T Ichiba ◽  
A Hachimori
Keyword(s):  
Thermophilic Bacterium ◽  
Complete Loss ◽  
Inorganic Pyrophosphatase ◽  
Histidine Residue ◽  
First Order ◽  
Diethyl Pyrocarbonate ◽  
Lysyl Endopeptidase ◽  
First Order Kinetics ◽  
Pseudo First Order

Treatment of the inorganic pyrophosphatase from thermophilic bacterium PS-3 with diethyl pyrocarbonate resulted in the almost complete loss of its activity, which followed pseudo-first-order kinetics. The presence of Mg2+ prevented the inactivation. Enzyme inactivated with diethyl pyrocarbonate was re-activated by hydroxylamine. The inactivation parallelled the amount of modified histidine residue, and a plot of the activity remaining against the amount of modified histidine residue suggested that the modification of one of two histidine residues totally inactivated the enzyme. The site involved was found to be located in a single lysyl endopeptidase-digest peptide derived from the ethoxy[14C]carbonylated enzyme. Amino acid analysis and sequence analysis of the peptide revealed that it comprised residues 96-119 of the inorganic pyrophosphatase from thermophilic bacterium PS-3. These results, when compared with those reported for the Escherichia coli and yeast enzymes, imply that His-118 of the inorganic pyrophosphatase from thermophilic bacterium PS-3 is located near the Mg(2+)-binding site and thus affects the binding of Mg2+.

scholarly journals MITOCHONDRIAL PROTEIN SYNTHESIS IN HELA CELLS

1974 ◽  
Vol 60 (3) ◽  
pp. 755-763 ◽  
Author(s):  
Jonas B. Galper
Keyword(s):  
Protein Synthesis ◽  
Ethidium Bromide ◽  
Mitochondrial Protein ◽  
Specific Protein ◽  
Mitochondrial Proteins ◽  
Mitochondrial Protein Synthesis ◽  
Initiator Trna ◽  
Low Concentrations ◽  
Separate Protein

HeLa cell mitochondrial proteins have been shown to be the products of two separate protein-synthesizing systems; one, the general cellular mechanism, sensitive to inhibition by cycloheximide, the other, a specific mitochondrial system subject to inhibition by low concentrations of chloramphenicol (Galper, J. B., and J. E. Darnell. 1971. J. Mol. Biol 57:363). Preliminary data have suggested that a mitochondrial N-formyl-methionyl-tRNA (f-Met-tRNA) might be the initiator tRNA in the latter (Galper, J. B., and J. E. Darnell. 1969. Biochem. Biophys. Res. Commun. 34:205; 1971. J. Mol. Biol. 57:363). It is demonstrated here that the synthesis of these endogenous mitochondrial proteins is also subject to inhibition by ethidium bromide and decays with a half-life of 1½–2 h in cultures incubated with low concentrations of this dye. The role of formylated f-Met-tRNA as the initiator tRNA in the synthesis of mitochondrial proteins is supported by data from several experiments. The rates of ethidium bromide inhibition of both the charging of f-Met-tRNA and of the synthesis of mitochondrial proteins are strikingly similar. Inhibition by aminopterin of the formylation of f-Met-tRNA greatly depresses the rate of mitochondrial-specific protein synthesis. In the absence of the synthesis of these proteins, respiration, the levels of cytochromes a–a3 and b, and the number of mitochondrial cristae are decreased. The implications of these findings as they relate to mitochondrial biogenesis are discussed.

ONIOM and ab-initio calculations on the mechanism of uncatalyzed peptide bond formation

2012 ◽  
Vol 90 (6) ◽  
pp. 691-700 ◽  
Author(s):  
Hadieh Monajemi ◽  
Mohammad Noh Daud ◽  
Sharifuddin Mohd. Zain ◽  
Wan Ahmad Tajuddin Wan Abdullah
Keyword(s):  
Amino Acids ◽  
Bond Formation ◽  
Computational Cost ◽  
Peptide Bond ◽  
Peptide Bond Formation ◽  
Initiator Trna ◽  
Reduced Rate ◽  
Surface Scan ◽  
A Site

Finding a proper transition structure for the peptide bond formation process can lead one to a better understanding of the role of ribosome in catalyzing this reaction. Using computer simulations, we performed the potential energy surface scan on the ester bond dissociation of P-site aminoacyl-tRNA and the peptide bond formation of P-site and A-site amino acids. The full fragments of initiator tRNAimet and elongator tRNAphe are attached to both cognate and non-cognate amino acids as the P-site substrate. The A-site amino acid for all four calculations is methionine. We used ONIOM calculations to reduce the computational cost. Our study illustrates the reduced rate of peptide bond formation for misacylated tRNAimet in the absence of ribosomal bases. The misacylated elongator tRNAphe, however, did not show any difference in its PES compared with that for the phe-tRNAphe. This demonstrates the structural specification of initiator tRNAimet for the amino acids side chain.

scholarly journals Role of a 300-Kilodalton Nuclear Complex in the Maturation of Trypanosoma brucei Initiator Methionyl-tRNA

2004 ◽  
Vol 3 (4) ◽  
pp. 893-899 ◽  
Author(s):  
George K. Arhin ◽  
Shuiyuan Shen ◽  
Henriette Irmer ◽  
Elisabetta Ullu ◽  
Christian Tschudi
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Rna Interference ◽  
Trypanosoma Brucei ◽  
Unicellular Eukaryote ◽  
Down Regulation ◽  
Parasitic Protozoa ◽  
Initiator Trna ◽  
Nuclear Complex ◽  
Posttranscriptional Processing

ABSTRACT tRNAs are transcribed as precursors containing 5′ leader and 3′ extensions that are removed by a series of posttranscriptional processing reactions to yield functional mature tRNAs. Here, we examined the maturation pathway of tRNAMet in Trypanosoma brucei, an early divergent unicellular eukaryote. We identified an approximately 300-kDa complex located in the nucleus of T. brucei that is required for trimming the 5′ leader of initiator tRNAMet precursors. One of the subunits of the complex (T. brucei MT40 [TbMT40]) is a putative methyltransferase and a homolog of Saccharomyces cerevisiae Gcd14, which is essential for 1-methyladenosine modification in tRNAs. Down-regulation of TbMT40 by RNA interference resulted in the accumulation of precursor initiator tRNAMet containing 5′ extensions but processed 3′ ends. In addition, immunoprecipitations with anti-La antibodies revealed initiator tRNAMet molecules with 5′ and 3′ extensions in TbMT40-silenced cells, albeit at a much lower level. Interestingly, silencing of TbMT40, as well as of TbMT53, a second subunit of the complex, led to an increase in the levels of mature elongator tRNAMet. Taken together, our data provide a glance at the maturation of tRNAs in parasitic protozoa and suggest that at least for initiator tRNAMet, 3′ trimming precedes 5′ processing.

scholarly journals The Rnf complex is a Na+ coupled respiratory enzyme in a fermenting bacterium, Thermotoga maritima

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Martin Kuhns ◽  
Dragan Trifunović ◽  
Harald Huber ◽  
Volker Müller
Keyword(s):  
Electron Transport ◽  
Ion Transport ◽  
Cytoplasmic Membrane ◽  
Thermotoga Maritima ◽  
Physiological Role ◽  
Thermophilic Bacterium ◽  
Respiratory Enzyme ◽  
Membrane Bound ◽  
Reverse Electron Transport

Abstractrnf genes are widespread in bacteria and biochemical and genetic data are in line with the hypothesis that they encode a membrane-bound enzyme that oxidizes reduced ferredoxin and reduces NAD and vice versa, coupled to ion transport across the cytoplasmic membrane. The Rnf complex is of critical importance in many bacteria for energy conservation but also for reverse electron transport to drive ferredoxin reduction. However, the enzyme has never been purified and thus, ion transport could not be demonstrated yet. Here, we have purified the Rnf complex from the anaerobic, fermenting thermophilic bacterium Thermotoga maritima and show that is a primary Na+ pump. These studies provide the proof that the Rnf complex is indeed an ion (Na+) translocating, respiratory enzyme. Together with a Na+-F1FO ATP synthase it builds a simple, two-limb respiratory chain in T. maritima. The physiological role of electron transport phosphorylation in a fermenting bacterium is discussed.

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