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At least 307 records · Page 17Linked to original sources
PubMed1994

Time course of lipid peroxidation in puromycin aminonucleoside-induced nephropathy.

Reactive oxygen species have been implicated in the pathogenesis of acute puromycin aminonucleoside (PAN)-induced nephropathy, with antioxidants significantly reducing the proteinuria. The temporal relationship between lipid peroxidation in the kidney and proteinuria was examined in this study. Rats were treated with a single IV injection of puromycin aminonucleoside, (PAN, 7.5 mg/kg) and 24 hour urine samples were obtained prior to sacrifice on days 3,5,7,10,17,27,41 (N = 5-10 per group). The kidneys were removed, flushed with ice cold TRIS buffer. Kidney cortices from each animal were used to prepare homogenates. Tissue lipid peroxidation was measured in whole homogenates as well as in lipid extracts from homogenates as thiobarbituric acid reactive substances. Proteinuria was evident at day 5, peaked at day 7 and persisted to day 27. Lipid peroxidation in homogenates was maximal at day 3 and declined rapidly to control levels by day 17. This study supports the role of lipid peroxidation in mediating the proteinuric injury in PAN nephropathy.

Animals↗
PubMed1994

Analysis of the puromycin binding site in the 70 S ribosome of Escherichia coli at the peptide level.

Photoinduced cross-linking of Escherichia coli 70 S ribosomes with [3H]puromycin has led to the labeling of ribosomal proteins S7, S14, S18, L18, and L29. Proteolytic fragmentation of these proteins and separation of the peptide mixtures by C18 reversed-phase high performance liquid chromatography resulted in six puromycin-labeled peptides which were applied to sequence analysis. The following peptides were found labeled: Pro1-Lys10 of S7, Ala28-Lys46 and Ala7-Lys11 of S14, Asp24-Lys29 of S18, Tyr64-Lys68 of L18, and Thr55-Lys60 of L29. For the first time the molecular environment of an antibiotic in the procaryotic ribosome is presented at the peptide level.

Amino Acid Sequence↗
PubMed1975

[The effect of puromycin, nalidixic acid, and cycloheximide acetylcholinesterase purified from Electrophorus electricus L].

The action of puromycin, nalidixic acid and cycloheximide on acetylcholinesterase has been studied using an "electrophoretically pure" preparation. Results obtained show that these well known inhibitors of protein synthesis have an inhibitory effect on acetylcholinesterase activity. The inhibitory action exercised by puromycin and nalidixic acid is of a mixed inhibition type, similar to the one seen when decamethonium and d-tubocurarine are used as inhibitory agents. Cycloheximide acts in a non competitive way on the enzyme activity.

Cycloheximide↗
PubMed1975

[Effect of puromycin, nalidixic acid and cycloheximide on neuromuscular transmission].

The action of puromycin, nalidixic acid and cycloheximid on neuromuscular transmission has been assayed using diaphragm-phrenic nerve preparation from rat. The results demonstrate that all antibiotics used, reversibly blodk neuromuscular transmission. Cycloheximid seems to act presynaptically by inhibiting acetylcholine release from nerve terminal. Puromycin and nalidixic acid act as curare-like agents. They seem to compete with acetylcholine for fixation at the cholinergic receptor.

Animals↗
PubMed1977

A scanning and transmission electron microscopic comparison of puromycin aminonucleoside-induced nephrosis to hyperalbuminemia-induced proteinuria with emphasis on kidney podocyte pedicel loss.

Ultrastructural changes in the visceral epithelium and proximal tubules of rats were studied by scanning and transmission electron microscopy during the onset and progression of puromycin aminonucleoside nephrosis (PAN)-induced proteinuria. These changes were compared with those that occur during a similar degree of proteinuria induced by intraperitoneal injections of albumin. With the onset of proteinuria and oliguria, PAN rats exhibit loss of podocyte pedicels and podocyte major processes, an increase in pinocytotic activity, and an accumulation of cytoplasmic vacuoles and granules of variable size, shape, and electron density. Loss of podocyte pedicels involves a gradual decrease in pedicel height beginning at the pedicel tip and progressing down the pedicel arm, formation of nublike protrusions and interpedicel microbridges (35 to 45 nm. in width and 40 to 60 nm. in length) along the pedicel's base, the merging of microbridges to form more extensive regions of interpedicel contact, and a gradual broadening and retraction of pedicels. In response to hyperalbuminemia-induced proteinuria, kidney podocytes exhibit reactions during PAN, however, the podocyte pedicels, slit pores, and major processes of rats with hyperalbuminemia-induced proteinuria remain discrete. The loss of pedicels and major processes during PAN, therefore, apparently results from the effects of puromycin aminonucleoside per se rather than from the proteinuria associated with this disease. The proximal tubules of rats with hyperalbuminemia-induced proteinurea exhibit the same characteristic changes as PAN rat proximal tubules (i.e., loss of brush border, dilated lumina, abnormally thin walls, and accumulation of periodic acid-Schiff positive electron-dense luminal casts and cytoplasmic protein absorption droplets). The significance of these ultrastructural findings during PAN and hyperalbuminemia-induced proteinurea are discussed in terms of the etiology of PAN.

Albumins↗
PubMed1977

The effects of actinomycin D and puromycin on the induction of the cytosol enzymes involved in the androgen metabolism by testosterone in female rat liver.

Induction of the cytosol enzymes involved in the androgen metabolism in the female rat liver by testosterone was studied. The treatment by testosterone propionate increased the production of 5beta-androstane-3alpha,17beta-diol in the 105.000xg supernatant when 4-14C-testosterone was used as a substrate. The conversion of 4-14C-androstenedione to labelled testosterone was increased by the injection of testosterone propionate into female rats. Actinomycin D or puromycin which was administered for four days with daily subcutaneous injections of testosterone propionate prevented the increase of these enzymes by testosterone propionate. Actinomycin D given once with the final injection of testosterone propionate blocked the induction of cytosol 17beta-hydroxysteroid dehydrogenase by the treatment of testosterone propionate, whereas puromycin given in the same manner as actinomycin D had no such a blocking effect on the enzyme. Both of these antibiotics given once with the final injection of testosterone propionate showed no preventing effects on the induction of cytosol delta4-5beta-reductase by testosterone propionate.

Androgens↗
PubMed1985

Cloning and expression of a puromycin N-acetyl transferase gene from Streptomyces alboniger in Streptomyces lividans and Escherichia coli.

A gene (pac) encoding a puromycin N-acetyl transferase (PAC) of Streptomyces alboniger ATCC12461 was cloned in the Streptomyces plasmid pIJ702 and expressed in S. lividans 1326. Several clones resistant to puromycin were isolated and shown to carry pIJ702 with different inserts of S. alboniger DNA. They were classified as of low and high activity according to the levels of enzymatic activity expressed by them. The different levels of expression were related to the two possible orientations of the S. alboniger DNA inserts in the pIJ702 vector. Six of the recombinant plasmids contain a common 1.6-kb DNA sequence which, by subcloning experiments, was shown to carry a pac gene encoding PAC activity. The pac gene was subcloned next to the lac promoter of Escherichia coli plasmid pUC19. Only one of the two possible orientations of insertion expressed PAC activity, suggesting that it was dependent on the lac promoter. Accordingly, isopropylthio-beta-D-galactoside (IPTG) was able to stimulate the expression of the enzyme activity. These results allowed the direction of transcription of the pac gene to be determined.

Acetyltransferases↗
PubMed1991

Molecular analysis of the dmpM gene encoding an O-demethyl puromycin O-methyltransferase from Streptomyces alboniger.

The nucleotide (nt) sequence of a 1332-bp fragment of Streptomyces alboniger DNA containing the gene (dmpM), which encodes an O-demethyl puromycin O-methyltransferase (DMPM), has been determined. The dmpM gene contains a 1131-nt open reading frame which encodes a polypeptide of Mr 40,303; this is consistent with the 44 +/- 2.5- and 160-kDa sizes of the DMPM monomer and its native form, respectively. The ATG start codon of dmpM is 50 bp downstream from the coding sequence of the gene (pac), which determines a puromycin N-acetyltransferase. S1 mapping experiments indicate that pac and dmpM are transcribed on a single transcript, which ends at least 500 nt downstream from the dmpM stop codon. The deduced amino acid sequence of DMPM shows significant similarities to those of a hydroxyindole O-methyltransferase, which is involved in the biosynthesis of melatonin by bovine pineal glands [Ishida et al., J. Biol. Chem. 262 (1987) 2895-2899], a hydroxyneurosporene methyltransferase, which is involved in carotenoid biosynthesis in the purple nonsulfur bacterium, Rhodobacter capsulatus [Armstrong et al., Mol. Gen. Genet. 216 (1989) 254-268] and two O-methyltransferases of the tetracenomycin biosynthesis pathway from Streptomyces glaucescens.

Acetylserotonin O-Methyltransferase↗
PubMed2003

Neuron-specific aminopeptidase and puromycin-sensitive aminopeptidase in rat brain development.

Neuron-specific aminopeptidase (NAP) and the ubiquitous puromycin-sensitive aminopeptidase (PSA) were compared in the rat hippocampus during early development. Hippocampus contains the highest amount of NAP determined by a fast-protein liquid chromatography-aminopeptidase analyzer using Leu beta-naphthylamide as substrate. Both enzymes were found in the hippocampus in all ages. NAP was lower in immature rat; the 19th embryonic-day fetus contained the least. It increased steeply during the prenatal through the early postnatal period, 9-fold by the first month. The rate of increase diminished subsequently, increasing 20% in the second month and 13% in the third. The age-dependent increase in NAP activity was parallel to its protein expression as determined by Western blot. The specific molecular activity (hydrolytic activity/ NAP antigenicity) in newborn, 15-day-old, and 30-day-old rats were 1.00, 0.88, and 1.00, respectively. The PSA developmental profile without linear increase in activity was distinct from NAP. PSA activity was higher than NAP in decreasing order, 100-4 times, during the same development span. Similarly, different growth profiles for NAP and PSA were also found in the primary culture of developing cerebellar granule cells. Puromycin (1-5 microM) blocked neurite outgrowth and caused apoptosis by nonantibiotic effects. Our data suggest that the synaptosome-enriched NAP plays a role in neuron growth, differentiation, and information programming.

Aging↗
PubMed1965

Accumulation of amino acids in muscle of perfused rat heart. Effect of insulin in the presence of puromycin.

1. Rat heart was perfused with a mixture of amino acids and other ninhydrin-positive substances in the same concentrations as they are found in plasma, and with or without sufficient puromycin to suppress protein synthesis all but completely. 2. In the presence of puromycin, but not in its absence, insulin increased the concentrations of most of the individual amino acids in the intracellular water of heart. 3. The results accord with the conclusion that insulin increases the entry of most, if not all, amino acids into muscle.

Journal Article↗
PubMed1971

Dissociation of mammalian polyribosomes into subunits by puromycin.

Hepatic ribosomes have been dissociated into biologically active subunits as follows. Polysomes were treated at 0 degrees C with puromycin at high ionic strength. This released most of the nascent polypeptide chains without dissociating the polysomes, which retained the mRNA and the tRNA moiety of peptidyl tRNA, but were unable to continue the translation of mRNA. The polysomes were then heated to 37 degrees C, when they dissociated completely into subunits. Similar treatment without puromycin resulted in only partial dissociation.

Amino Acids↗
PubMed2006

The puromycin-sensitive aminopeptidase PAM-1 is required for meiotic exit and anteroposterior polarity in the one-cell Caenorhabditis elegans embryo.

In the nematode Caenorhabditis elegans, sperm entry into the oocyte triggers the completion of meiosis and the establishment of the embryonic anteroposterior (AP) axis. How the early embryo makes the transition from a meiotic to a mitotic zygote and coordinates cell cycle changes with axis formation remains unclear. We have discovered roles for the C. elegans puromycin-sensitive aminopeptidase PAM-1 in both cell cycle progression and AP axis formation, further implicating proteolytic regulation in these processes. pam-1 mutant embryos exhibit a delay in exit from meiosis: thus, this peptidase is required for progression to mitotic interphase. In addition, the centrosomes associated with the sperm pronucleus fail to closely associate with the posterior cortex in pam-1 mutants, and the AP axis is not specified. The meiotic exit and polarity defects are separable, as inactivation of the B-type cyclin CYB-3 in pam-1 mutants rescues the meiotic exit delay but not the polarity defects. Thus PAM-1 may regulate CYB-3 during meiotic exit but presumably targets other protein(s) to regulate polarity. We also show that the pam-1 gene is expressed both maternally and paternally, providing additional evidence that sperm-donated gene products have important roles during early embryogenesis in C. elegans. The degradation of proteins through ubiquitin-mediated proteolysis has been previously shown to regulate the cell cycle and AP axis formation in the C. elegans zygote. Our analysis of PAM-1 requirements shows that a puromycin-sensitive aminopeptidase is also required for proteolytic regulation of the oocyte to embryo transition.

Amino Acid Sequence↗
PubMed1979

Selective inhibition of preribosomal RNA synthesis by puromycin aminonucleoside in transformed human fibroblasts: studies of the nature of the inhibition in isolated nuclei and nucleoli.

Puromycin aminonucleoside selectively inhibits the synthesis of ribosomal RNA in human lung fibroblasts transformed by the oncogenic virus SV40. The mechanism of this inhibition was studied utilizing nuclei and nucleoli isolated from cells treated for 18 hours with 100 micrograms/ml of this compound. It was established that for a limited period of time nuclei and nucleoli isolated from the fibroblasts continue synthesis of RNA of size classes seen in intact cells, and that the inhibitory effect of aminonucleoside persists after isolation of these organelles. The inhibition was shown to be directed primarily to the activity of RNA polymerase I. Studies of the mechanism of this inhibition have indicated that the decreased rate of the polymerase reaction is not due to the impairment of the template function of nucleolar chromatin, and that unbound, as well as chromatin-bound, RNA polymerase I is present in both control and treated nucleoli. Analysis of the size distribution of the products of cell-free RNA synthesis showed that aminonucleoside pretreatment results in marked reduction in the synthesis of preribosomal 45S RNA, abnormal accumulation of 32S RNA, and reduced formation of mature ribosomal RNA species in the in vitro system. The data suggest that the inhibitory effect of aminonucleoside on ribosomal synthesis is due in part to a lower rate of transcription by RNA polymerase I of preribosomal RNA, and in part to its impaired maturation.

Cell Line↗
PubMed1987

DMSO potentiates aminonucleoside of puromycin nephrosis in rats.

Dimethyl sulphoxide (DMSO), 3 g/kg body weight, administered daily by the intraperitoneal route, potentiated the proteinuria and formation of tubular casts in aminonucleoside of puromycin (PA) induced nephrosis in Sprague-Dawley rats. The effect was evident at 4 as well as 8-9 days following PA administration. In the absence of PA, DMSO did not induce proteinuria or cast formation. The mechanism by which DMSO enhanced proteinuria and cast formation is not known.

Animals↗
PubMed1990

In vitro effects of puromycin aminonucleoside on the ultrastructure of rat glomerular podocytes.

Puromycin aminonucleoside (PAN)-induced nephrosis in rats provides a model for studying the pathogenesis of severe proteinuric conditions, such as minimal change disease. The present study used scanning (SEM) and transmission (TEM) electron microscopy to investigate the in vitro effects of PAN on rat glomerular podocytes. Slices of rat kidney were incubated for up to 3 days in Medium 199 with Hanks' salts (control) or in medium with PAN. Semiquantitative SEM analysis of glomeruli on the upper surface of kidney slices indicated that incubation with PAN (100 micrograms/ml and 500 micrograms/ml) decreased the number of microvilli on podocyte cell bodies (days 1, 2 and 3), increased the number of glomeruli showing flattening of podocyte cell bodies and major processes (days 2 and 3), and increased the number of glomeruli showing surface membrane blebbing on podocyte foot processes (day 3) (p less than 0.001 in all cases). TEM morphometry revealed that incubation with 500 micrograms/ml PAN retarded significantly (p less than 0.001 at days 2 and 3) the loss of podocyte foot processes observed in control cultures. Whilst the SEM changes to podocyte ultrastructure largely mimic those seen in PAN nephrosis in vivo, the retardation of foot process loss runs counter to the major TEM change observed in vivo.

Animals↗
PubMed1979

Inhibition of cellular transition from G1-resting to G1-prereplicative phase by aminonucleoside of puromycin.

Human embryonic lung fibroblasts (IMR-90 and WI-38) were arrested in the G1 phase of the cell cycle by serum deprivation and high population density. Within 1 hr after the addition of medium containing fresh serum, these cells showed an increase in rRNA synthesis. The inclusion of 100 micrograms per ml aminonucleoside of puromycin (AMS) in the fresh medium eliminated the serum stimulation of rRNA synthesis and prevented the cells from making the G1-resting phase to G1-prereplicative phase transition. AMS also prevented the synthesis of HnRNA normally found within 10 hr after serum stimulation. Serum-stimulated RNA synthesis in starved, SV-40 transformed fibroblasts (WI-38-VA-13 cells) was inhibited, but not completely prevented, by AMS indicating that transformed cells may produce specific RNA's that are not AMS-sensitive and that may be responsible for the failure of transformed cells to be arrested in G1.

Animals↗
PubMed1983

A study on cadmium-induced nephropathy in rats pretreated with puromycin aminonucleoside.

Nephrotoxicity of cadmium (Cd) was investigated using puromycin aminonucleoside (AN)-pretreated rats. AN pretreatment was performed by iv injection of 100 mg AN/kg body wt 11 days before the initial Cd injection. Since massive proteinuria and focal glomerular deposits were recognized, glomerular permeability is considered to be increased in AN-pretreated rats. AN-pretreated and intact rats were injected sc with 3 mg Cd/kg body wt, 4 times a week. In non-pretreated rats, slight tubular vacuolation was seen after 1-week Cd exposure and severe vacuolation and coagulative necrosis of the tubules was observed after 2-week Cd exposure. On the other hand, AN pretreatment delayed the onset of vacuolation and necrosis for 1 week and made the lesion milder. After 1-week Cd exposure, a larger amount of Cd was excreted into the urine of AN-pretreated rats than of non-pretreated ones, whereas Cd accumulation in the kidney and liver was lower in AN-pretreated rats than in non-pretreated ones. Thereafter, no difference in Cd concentration was recognized between two groups. From these findings, it is suggested that in early stage of Cd administration, Cd was filtered through the glomerular basement membrane modified by AN pretreatment and that this filterable Cd did not have nephrotoxic effects in AN-pretreated rats.

Animals↗
PubMed1976

Intracellular distribution of 8-14C-puromycin aminonucleoside in ultraviolet irradiated Escherichia coli.

The uptake of 8-14C-puromycin aminonucleoside (8-14C-PAN) was studied in ultraviolet (UV) irradiated strains of E. coli B/r hcr+ and hcr-. The cells took up only 0.1-0.3% of the 8-14C-PAN present in the medium when grown in minimal (M9) containing 8-14C-PAN. When ethylenedinitrilotetra-acetic acid (EDTA) treated E. coli cells are placed in a medium containing 8-14C-PAN, the total concentration of 8-14C-PAN in the cells reaches 43-54% of the medium within 30 min of incubation. Almost all 8-14C-PAN can be dialyzed from cells exposed in the absence of an energy source, but cells metabolizing in M9 medium during exposure can retain up to 30% of their internal concentration. Bacteria grown in the presence of 8-14C-PAN, accumulated the radioactive material intracellularly in three forms, namely, unbound, reversibly bound (dialyzable) and irreversibly bound to the protein (nondialyzable). Approx. 70-77% of the irreversibly bound radioactive material linked with the protein fraction was released by treatment with a protease. Addition of PAN into the post-irradiation medium of EDTA-treated hcr+ cells, increased UV induced mutation rates. Antimutagenic purine ribosides decreased the final level of 8-14C-PAN accumulated by the cells. Decreases in 8-14C-PAN uptake in the presence of antimutagens correspond to reductions in the rate of mutation to streptomycin resistance induced by UV light. Therefore, protein bound PAN appears to be the relevant component in the enhancement of UV induced mutation by this drug.

Biological Transport↗