Two novel mitochondrial and chloroplastic targeting-peptide-degrading peptidasomes in A. thaliana, AtPreP1 and AtPreP2

Two novel metalloendopeptidases in Arabidopsis thaliana, AtPreP1 and AtPreP2, are responsible for the degradation of targeting peptides in mitochondria and chloroplasts. Both AtPreP1 and AtPreP2 contain ambiguous targeting peptides and are dually targeted to both organelles. The proteases also have the capacity to degrade unstructured peptides of up to 65 amino acid residues, but not small proteins. The catalysis occurs in a huge catalytic chamber revealed by the crystal structure of AtPreP1 at 2.1 A. The enzymes show a preference for basic and small uncharged amino acids or serines at the cleavage sites. Despite similarities in cleavage specificities, cleavage-site recognition differs for both proteases and is context- and structure-dependent. The AtPreP1 and AtPreP2 genes are differentially expressed in Arabidopsis.     

The precursor of the F1β subunit of the ATP synthase is covalently modified upon binding to plant mitochondria

We present evidence for a unique covalent modification of a nuclear-encoded precursor protein targeted to plant mitochondria. We investigated the early events of in vitro import for the mitochondrial precursor of the ATP synthase F₁ subunit from Nicotiana plumbaginifolia (pF₁) into plant mitochondria. When pF₁ of 59 kDa was incubated with mitochondria isolated from different higher-plant species, a band of 61 kDa was generated. The 61 kDa protein was a covalently modified form of the 59 kDa pF₁. The modification was dependent on the 25 amino acid long N-terminal region of the presequence of pF₁. The modification was catalysed by an enzyme located in the outer mitochondrial membrane which was specific for higher plants and could not be washed off from the membrane by urea, KCl or EDTA. The modification was ATP- and Ca²⁺-dependent, but it was not affected by inhibitors of protein kinases. No inhibition of the modification was observed with phosphatase, methylation or acylation inhibitors. The modification occurs prior to translocation through the mitochondrial outer membrane. Inhibition of the modification process does not affect the import of the precursor protein, hence precursor modification was not a prerequisite for import. Both the modified and the unmodified pF₁ proteins were strongly associated with the mitochondrial outer membrane.     

Plasma levels of total, free and protein bound thiols as well as sulfane sulfur in different age groups of rats

The redox status of plasma thiols can be a diagnostic indicator of different pathological states. The aim of this study was to identify the age dependent changes in the plasma levels of total, free and protein bound glutathione, cysteine and homocysteine. The determination was conducted in plasma of three groups of rats: 1) young (3-month-old), 2) middle aged (19-month-old), and 3) old (31-month-old). Total levels of glutathione, cysteine and homocysteine and their respective free and protein-bound fractions decreased with age. The only exception was a rise in free homocysteine concentration in the middle group, which indicates a different pattern of transformations of this thiol in plasma. The drop in the level of protein-bound thiols suggests that the antioxidant capacity of plasma diminishes with age, which, consequently, leads to impaired protection of -SH groups through irreversible oxidation. The plasma sulfane sulfur level also declines with age, which means that aging is accompanied by inhibition of anaerobic sulfur metabolism.     

Experimental hybridisation between X0 and XY chromosome races in the grasshopper Podisma sapporensis Shir. (Orthoptera, Acrididae). I. Cytological analysis of embryos and F1 hybrids

The results of experimental hybridisation between some chromosome subraces belonging to the X0 and XY chromosome races of the brachypterous grasshopper P. sapporensis are presented. Pre-zygotic reproductive isolation mechanisms in experimental pairs were not confirmed. In crossings of XY-standard x X0-standard and XY-standard x X0-Naganuma chromosome subraces, a zygotic barrier has been found. All embryos of XY-standard x X0-standard crosses and the vast majority of embryos of XY-standard x X0-Naganuma crosses were obtained from female diploid or haploid/diploid cells as a result of parthenogenesis. In very rare cases, when the zygotic barriers had been surmounted, normal embryo heterozygotes and a F1 hybrid generation were obtained in XY-standard x X0-Naganuma crosses. On the contrary, crosses between the XY-Tanno and X0-standard subraces gave viable offspring in spite of many chromosome differences such as a X-A translocation and fixed pericentric inversions in four pairs of autosomes. The results obtained do not support the hypothesis that chromosomal differences play a key role in restricting gene flow between X0 and XY races of P. sapporensis. The presence of crossing barriers explains the phenomena of the purity of the X0 and XY chromosomes races.     

Instructions for authors

The aim of the present study was to examine hypothesis that the enhanced cholesterologenesis, found in rats with experimental chronic renal failure (CRF) resulted from the increased gene expression of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase – the rate limiting enzyme in the cholesterologenesis pathway, responsible for mevalonate synthesis. Wistar rats were used and experimental CRF was achieved by 5/6 nephrectomy model. We examined: (a) the changes in the rat liver microsomal HMG-CoA reductase activity, (b) the rat liver HMG-CoA reductase mRNA abundance in various times of day. Obtained data indicates that the increased activity of HMG-CoA reductase in the liver of rats with experimental CRF parallel enhanced mRNA level and suggests that enhanced cholesterol biosynthesis, observed in experimental CRF is at least in part due to the increased HMG-CoA reductase gene expression. The results also indicate that the physiological diurnal rhythm of HMG-CoA reductase activity is preserved in the course of experimental CRF.     

Regulation of c-Jun N-terminal kinase,p38 kinase and AP-1 DNA binding in cultured brain neurons: roles in glutamate excitotoxicity and lithium neuroprotection

In rat cerebellar granule cells, glutamate induced rapid activation of c-Jun N-terminal kinase (JNK) and p38 kinase to phosphorylate c-Jun (at Ser63) and p53 (at Ser15), respectively, and a subsequent marked increase in activator protein-1 (AP-1) binding that preceded apoptotic death. These glutamate-induced effects and apoptosis could largely be prevented by long-term (7 days) pretreatment with 0.5-2 mm lithium, an antibipolar drug. Glutamate's actions could also be prevented by known blockers of this pathway, MK-801 (an NMDA receptor blocker), SB 203580 (a p38 kinase inhibitor) and curcumin (an AP-1 binding inhibitor). The concentration- and time-dependent suppression of glutamate's effects by lithium and curcumin correlated well with their neuroprotective effects. These results suggest a prominent role of JNK and p38, as well as their downstream AP-1 binding activation and p53 phosphorylation in mediating glutamate excitotoxicity. Moreover, the neuroprotective effects of lithium are mediated, at least in part, by suppressing NMDA receptor-mediated activation of the mitogen-activated protein kinase pathway.     

Induction of apoptosis by EGCG in selected tumour cell lines in vitro

One of the best recognised polyphenols of plant origin, epigallocatechin-3-gallate (EGCG) is contained mainly in green tea and in grapes. Studies performed in vivo and in vitro have demonstrated high probability of anti-neoplastic potential of the compound, due to its capacity to induce programmed cell death. The present studies were aimed at evaluation of apoptosis induction in cells of three selected tumour cell lines, subjected to action of various concentrations of EGCG. The experiment was performed on cultures of HEp-2 laryngeal carcinoma cells, LoVo colon carcinoma cells, HeLa cervical carcinoma cells and on normal myoepithelial cell line, HS. EGCG was found to induce apoptosis in cells of the examined neoplastic lines in a dose-related manner. Moreover, effect of EGCG on normal cells of HS line was found to be much less pronounced as compared to effects exerted on sensitive neoplastic cells.