Properties of liver mitochondria from iron-loaded rats.

The increased iron content in livers from iron-loaded rats is almost exclusively confined to the mitochondria. The ten- to twenty-fold higher level of nonheme iron in such mitochondria decreases the respiratory control with pyruvate-malate, but not with 3-hydroxybutyrate or succinate as substrates, and has no effect on the capacity for phosphorylation and substrate oxidation. Iron-loaded mitochondria have a malondialdehyde level which is about three times higher than that of control mitochondria, even after repeated washings with bovine serum albumin and EDTA. This is suggestive of an on-going process of lipid oxidation presumably catalyzed by the accumulated iron. Differences between the present in vivo data and in vitro results obtained by others are discussed.     

Some characteristics of mitochondrial multienzyme systems from rat liver mitochondria after heating of rats]

A character of rat liver mitochondria degradation after the heat treatment of animals is studied. It is found that mitochondria under the effect of elevated temperature do not considerably change their functional characteristics and thus they are capable to provide the normal rate of ATP synthesis, the rate of succinate oxidation being slightly increased. At the same time the heating caused the degradation of mitochondria which results in the decrease of their thermostability, in the increased susceptibility to lytic effect of trypsin and phospholipase D, and in the activation of succinate dehydrogenase and cytochrome c oxidase. The mitochondria degradation is due to the formation of "latent impairments" in the structure of mitochondria.     

Transmembrane potential of liver mitochondria from hexachlorobenzene-and iron-treated rats

The respiratory parameters and the membrane potential of liver mitochondria from rats treated with either hexachlorobenzene, iron or hexachlorobenzene plus iron, to induce experimental porphyria, have been studied. Partial uncoupling of oxidative phosphorylation has been observed in mitochondria from hexachlorobenzene- and hexachlorobenzene plus iron-treated rats. Direct evidence has been presented that this uncoupling is due to the action of pentachlorophenol endogenously formed by metabolism of hexachlorobenzene. No irreversible damage of mitochondria membrane has been revealed under both these conditions. Normal oxidative phosphorylation has been found in mitochondria from rats treated with iron alone. In contrast, they presented an anomalous membrane potential, fully restored by oligomycin. A possible involvement of lipid peroxidation process, induced by iron, in causing these abnormalities has been suggested.     

Protein kinases from liver mitochondria of tumour-bearing rats.

The mitochondria of liver of Yoshida ascites tumour-bearing rats contained two forms of protein kinase distinguishable on the basis of their kinetic properties, substrate specificity and responses to cyclic adenosine 3',5'-monophosphate (cAMP). One of these (kinase I) was activated 2-3 fold by cAMP while the other form (kinase II) was insensitive to the action of cAMP. Kinase I which was selective towards histone F1 as substrate was obtained as a homogeneous preparation and was observed to have a molecular weight of 170 000 by Sephadex G-150 gel filtration. Protein kinase II appeared to be a smaller protein with molecular weight of 54 000 and was specific towards acidic proteins namely casein and phosvitin. Protein kinases isolated from liver mitochondria of normal rats showed variations in respect to elution profile of DEAE-cellulose and electrophoretic mobility. The preparation corresponding to kinase I did not show stimulatory responses to cAMP.     

Various properties of ribonucleotide reductase from rat liver mitochondria]

Three fractions of mitochondria (light, heavy and intermediate) from normal and regenerating rat liver ( in 15, 24 and 48 hours after partial hepatectomy) were isolated by means of differential centrifugation. Heavy mitochondrial fraction was shown to have a higher CDP-reductase activity than light mitochondria. The activity of the procces of ribonucleotides reduction in the mitochondria was shown to depend on the tissue functional state and it was maximal in 24 hours after partial hepatectomy.     

Transmembrane potential of liver mitochondria from hexachlorobenzene- and iron-treated rats

The respiratory parameters and the membrane of liver mitochondria from rats treated with either hexachlorobenzene, iron or hexachlorobenzene plus iron, to induce experimental porphyria, have been studied. Partial uncoupling of oxidative phosphorylation has been observed in mitochondria from hexachlorobenzen- and hexachlorobenzene plus iron-treated rats. Direct evidence has been pressented that this uncoupling is due to the action of pentochlorophenol endogenously formed by metabolism of hexachlorobenzene. No irreversible damage of mitochondrial membrane has been revealed under both these conditions. Normal oxidative phosphorylation has bee found in mitochondria from rats treated with iron alone. In contrast, they presented an anomalous membrane potential, fully restored by oligomycin. A possible involvement of lipid peroxidation process, induced by iron, in causing these abnormalities has been suggested.     

Sequences of three transfer RNAs from mosquito mitochondria

The sequences of three transfer RNAs from mosquito cell mitochondria, tRNA_UCG^Arg, tRNA_GUC^Asp, and tRNA_GAU^Ile, determined using a combination of rapid ladder and fingerprinting procedures are reported. These were compared with hamster mitochondrial tRNA_UCG^Arg and tRNA_GUC^Asp determined similarly, and a bovine mitochondrial tRNA_GAU^Ile determined using a somewhat different approach. The primary sequences of the mosquito tRNAs were 35 to 65% homologous to the corresponding mammalian mitochondrial species, and bore little homology to “conventional” (bacterial or eucaryotic cytoplasmic) tRNA. The modification status of the mosquito mitochondrial tRNAs resembled that of mammalian mitochondrial tRNA. The results contribute to the generalization that metazoan mitochondrial tRNA constitutes a distinctive, albeit loosely structured, phylogenetic group.     

Ion transport in liver mitochondria from normal and thyroxine-treated rats

Liver mitochondria isolated from rats 24 h after a single subcutaneous injection of 8 mg thyroxine per kilogram body weight were compared with those isolated from control rats that received injections of isotonic saline at the same time. The mitochondria isolated from the thyroxine-treated rats show higher rates of energy-dependent K⁺ and phosphate accumulation than those from control animals. It was also found that mitochondria from the hormone-treated animals required a larger addition of Ca²⁺/mg mitochondrial protein in order to uncouple oxidative phosphorylation, and showed smaller tendency to swellin vitro under energizing conditions. The data obtained on ion movements support previous observations that the stimulation of the basal rate of mitochondrial respiration by thyroxine is associated with an increase in the transmembrane protonic electrochemical potential difference, and indicate thatin vivo the hormone raises the intramitochondrial concentration of K⁺ and phosphate.     

The electrophoretic behaviour of mitochondria from kidney and liver of rats

1. The electrophoretic mobility values for mitochondria prepared from rat kidney and liver measured in 0.125m-potassium chloride-0.02m-tris, pH7.4, are: -0.78(s.e.m.+/-0.02)mu/sec./v/cm. (29 experiments) and -1.06(s.e.m.+/-0.01) mu/sec./v/cm. (21 experiments) respectively. 2. These mobility values are unaffected by washing and spontaneous swelling at 25 degrees , indicating a stable electrokinetic surface. 3. The mobility of rat-kidney mitochondria is unaffected by thyroxine-induced swelling, or by the state of hydration of the rat. 4. pH-mobility curves show similar surface ionogenic groups for kidney and liver mitochondria; their isoelectric points are pH3.9 and pH4.4 respectively.     

Ubiquinone content in albino rat kidney mitochondria following unilateral nephrectomy]

Following unilateral nephrectomy (in 72 hours) the weight of the remaining kidney increased by 23%, and the activity of succinic dehydrogenase--by 43.2%. The concentration of oubichinon in the hypertrophied kidney and its mitochondria showed no significant change for 3 days after the nephrectomy.     

Identification of small non-coding RNAs from mitochondria and chloroplasts

Small non-protein-coding RNAs (ncRNAs) have been identified in a wide spectrum of organisms ranging from bacteria to humans. In eukarya, systematic searches for ncRNAs have so far been restricted to the nuclear or cytosolic compartments of cells. Whether or not small stable non-coding RNA species also exist in cell organelles, in addition to tRNAs or ribosomal RNAs, is unknown. We have thus generated cDNA libraries from size-selected mammalian mitochondrial RNA and plant chloroplast RNA and searched for small ncRNA species in these two types of DNA-containing cell organelles. In total, we have identified 18 novel candidates for organellar ncRNAs in these two cellular compartments and confirmed expression of six of them by northern blot analysis or RNase A protection assays. Most candidate ncRNA genes map to intergenic regions of the organellar genomes. As found previously in bacteria, the presumptive ancestors of present-day chloroplasts and mitochondria, we also observed examples of antisense ncRNAs that potentially could target organelle-encoded mRNAs. The structural features of the identified ncRNAs as well as their possible cellular functions are discussed. The absence from our libraries of abundant small RNA species that are not encoded by the organellar genomes suggests that the import of RNAs into cell organelles is of very limited significance or does not occur at all.