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PubMed2003

A Synechococcus sp. PCC 7942 mutant with a higher tolerance towards bentazone.

In this article we describe the partial characterization of a Synechococcus sp. PCC 7942 mutant Mu1 with an enhanced resistance towards the herbicide bentazone (3-isopropyl-1H-2,1,3-benzothiadiazine-4(3H)-one 2,2-dioxide). The mutant was derived from a random mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine (NSG) and exhibited superior growth rates, pigment content and overall photosynthetic activities under regular growth conditions compared to wild type. Whereas Synechococcus PCC 7942 wild type showed significant photoinhibition, especially in the presence of lincomycin, Mu1 was much more robust. A comparative analysis of the content of several photosynthesis-associated proteins revealed that Mu1 had an increased expression of PsbO on mRNA and protein level and that PsbO is tightly bound to Photosystem II, relative to wild type. This result was substantiated by mass spectrometer measurements of photosynthetic water oxidation revealing a higher stability and integrity of the water oxidizing complex in Mu1 cells grown under regular or calcium deficient conditions. Therefore, our results give rise to the possibility that the overexpression of PsbO in mutant Mu1 confers resistance to reactive oxygen species (ROS) formed as a consequence of bentazone binding to the acceptor side of PS II. In addition, we observed a significantly higher tolerance towards bentazone in iron depleted wild type cells, conditions under which the IdiA protein becomes expressed in highly elevated amounts. As we have previously shown, IdiA preferentially protects the acceptor site of PS II against oxidative stress, especially under iron limitation. Thus, it is likely that IdiA due to its topology interferes with bentazone binding or protects PS II against ROS generated in the presence of bentazone.

Journal Article↗
PubMed2003

The interaction of visible and UV-B light during photodamage and repair of Photosystem II.

In order to understand the mechanism of photodamage induced by solar radiation under natural conditions, we studied the interaction of visible and ultraviolet-B light in the inactivation and repair of the Photosystem II complex by using oxygen evolution and flash-induced chlorophyll fluorescence measurements. In isolated spinach thylakoids and Synechocystis 6803 cells, in which de novo protein synthesis is blocked by lincomycin, photodamage of Photosystem II by visible and UV-B light is characterized by linear semilogarithmic inactivation curves for both separate and combined illumination protocols. The extent of PS II inactivation obtained after combined illumination can be well simulated by assuming independent damaging events induced by visible and UV-B photons. In intact Synechocystis cells capable of protein repair, simultaneous illumination by visible and UV-B light impairs Photosystem II activity to a smaller extent than expected from the independent damaging events. This protective effect is pronounced at low visible light (130 muE m(-2) s(-1)), but becomes negligible at high intensities (1300 muE m(-2) s(-1)). Exposure of intact Synechocystis 6803 cells to direct sunlight leads to a rapid inactivation of PS II, accompanied by the accumulation of donor side inhibited centers. This phenomenon, which shows the impairment of the manganese cluster of water oxidation was not observed when the ultraviolet components of sunlight were filtered out. We conclude that visible and UV-B photons inactivate PS II via non-interacting mechanisms, which affect different target sites. In intact cells, the two spectral regions do interact, and results in synergistically enhanced protein repair capacity when UV-B radiation is accompanied by low intensity visible light, which provides protection against photodamage. However, this ameliorating effect becomes insignificant at high light intensities characteristic of direct sunlight.

Journal Article↗
PubMed2001

Enhanced photochemical light utilization and decreased chilling-induced photoinhibition of photosystem II in cotton overexpressing genes encoding chloroplast-targeted antioxidant enzymes.

The aim of this study was to determine whether increases in stromal superoxide dismutase (SOD; EC 1.15.1.1), ascorbate peroxidase (APX; EC 1.11.1.11) and glutathione reductase (GR; EC 1.6.4.2) via transformation could reduce photosystem (PS) II photoinhibition at low temperature for cotton (Gossypium hirsutum L.) plants and to determine by what mechanism this protection may be realized. During 3-h exposures of lincomycin-treated leaf discs to 10 degrees C and a photon flux density of 500 &mgr;mol m-2 s-1, all transgenic plants exhibited significantly greater PSII activity and O2 evolution than did wild-type plants. Also, the rate constant of PSII photoinactivation was significantly lower for all transgenic plants than for wild-type plants. No significant differences existed between genotypes in non-photochemical quenching of chlorophyll a fluorescence and the regulated component of the thermal dissipation of excitation energy. The relationship between changes in variable to maximum chlorophyll fluorescence (Fv/Fm) and the time-dependent averaged excessive light exposure was similar for all genotypes. This observation excluded the possibility that differences in PSII photodamage were due to improvements in the direct protection of PSII from active oxygen by antioxidant enzyme overproduction. Similar decreases in Fv/Fm during the stress treatment for all genotypes when leaves were pre-treated with 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU) suggested that the effect of overproduction involved events downstream of PSII in the electron transfer pathway. Since all transgenic plants exhibited a significantly higher photochemical quenching of chlorophyll fluorescence during the chilling treatment, we concluded that, under the conditions used in this study, the enhancement of the protection of PSII from photodamage by increasing the stromal antioxidant enzyme activity in cotton leaves was due to the maintenance of a higher rate of electron transport and, consequently, a lower reduction state of QA.

Journal Article↗
PubMed2002

Photoinhibition and loss of photosystem II reaction centre proteins during senescence of soybean leaves. Enhancement of photoinhibition by the 'stay-green' mutation cytG.

The 'stay-green' mutation cytG in soybean (Glycine max) partially inhibits the degradation of the light-harvesting complex II (LHCII) and the associated chlorophyll during monocarpic senescence. cytG did not alter the breakdown of the cytochrome b6/f complex, thylakoid ATP synthase or components of Photosystem I. In contrast, cytG accelerated the loss of oxygen evolution activity and PSII reaction-centre proteins. These data suggest that LHCII and other thylakoid components are degraded by separate pathways. In leaves induced to senesce by darkness, cytG inhibited the breakdown of LHCII and chlorophyll, but it did not enhance the loss of PSII-core components, indicating that the accelerated degradation of PSII reaction centre proteins in cytG was light dependent. Illumination of mature and senescent leaves of wild-type soybean in the presence of an inhibitor (lincomycin) of chloroplast protein synthesis revealed that senescence per se did not affect the rate of photoinhibition in leaves. Likewise, mature leaves of the cytG mutant did not show more photoinhibition than wild-type leaves. However, in senescent cytG leaves, photoinhibition proceeded more rapidly than in the wild-type. We conclude that the cytG mutation enhances photoinhibition in senescing leaves, and photoinhibition causes the rapid loss of PSII reaction-centre proteins during senescence in cytG.

Journal Article↗
PubMed1985

A conjugative plasmid encoding production of a diffusible pigment and resistance to aminoglycosides and macrolides in Staphylococcus aureus.

A methicillin-resistant strain of Staphylococcus aureus, WG512, was isolated in 1968 and has been shown to harbour a plasmid, pWG14, which encodes the production of a diffusible pigment and resistance to kanamycin, neomycin, streptomycin, lincomycin, erythromycin and spectinomycin. Plasmid pWG14 has only been detected as a slowly migrating band of DNA during agarose-gel electrophoresis, and this was thought to be the open-circular form of the plasmid with an estimated molecular weight of 30-34 Mdal. The transfer properties of this plasmid were compared with a non-conjugative plasmid encoding tetracycline resistance and the class 2, conjugative plasmid described by Archer and Johnston (1983). Plasmid pWG14 shared many of the conjugative properties of the latter plasmid, including the ability to transfer at high frequency on the surface of a filter membrane and to transfer between non-lysogenic strains in the absence of Ca++ and in the presence of citrate of DNase I. However, unlike the class 2 plasmid, pWG14 was able to transfer in broth culture at low frequency and did not mobilise the non-conjugative plasmid. Furthermore, the class 2 plasmid and pWG14 were shown to belong to different incompatibility groups. A derivative of WG512, which had lost the ability to produce the diffusible pigment along with plasmid pWG14 but had retained the resistance determinants of plasmid pWG14 in the chromosome, was used to demonstrate that the conjugative mechanism of pWG14 was a property of the plasmid and not the resistance determinants.

Aminoglycosides↗
PubMed1998

HPLC determination of residues of spectinomycin in various tissue types from husbandry animals.

An HPLC method was developed for the determination of bacteriostatic aminocyclitol spectinomycin (SP) in animal tissue products. These products included chicken eggs and edible fat, kidney, liver, muscle tissues from calf, poultry, pig and sheep. Residues of SP were extracted from homogenized tissue and egg-derived material with 25 mM citrate of pH 4.0, trichloroacetic acid and dichloromethane. The extract was purified and concentrated over a carboxylic acid-bonded solid-phase extraction (SPE) column. The SPE-eluate was analysed by cation-exchange HPLC involving a two-column switching system, post-column derivatization and fluorescence detection. Spectinomycin could be successfully determined at levels of 0.05 mg kg-1 and higher. Recoveries from spiked tissue material and from spiked egg material were in excess of 74% and did not show a concentration or tissue-type dependence. Precision of the elution position and signal response was better than 2%. Matrix effects and interference from lincomycin were less than 7 and 2%, respectively, on the signal response. Spectinomycin was shown to be stable at -20 degrees C in combined egg yolk and white over a test period of 12 weeks and in calf and sheep muscle tissue over a test period of 10 days. SP was, however, not stable at this temperature over a period of 12 months in chicken muscle tissue. Incurred SP residues were successfully determined in kidney and muscle tissue at the injection site of pigs administered with two doses of 15 mg kg-1 body weight SP with an intermittent withdrawal period of 15 days. Kidney showed higher concentrations and more persistent residues of SP than muscle tissue at the injection site.

Animals↗
PubMed1998

Detection of antibiotics in muscle tissue with microbiological inhibition tests: effects of the matrix.

The effects of the tissue matrix on detection limits of antibiotics with microbiological inhibition tests, intended for muscle tissue, were measured. Pieces of frozen meat were laid directly on top of paper disks impregnated with aqueous antibiotic solutions. Inhibition zones were compared with those obtained by the same standard solution without tissue. Only tetracyclines were detected as efficiently with as without muscle tissue. Inhibition zones of the beta-lactam antibiotics ampicillin and penicillin G, and the fluoroquinolone antibiotics enrofloxacin and ciprofloxacin were smaller when muscle tissue was added to low levels of standard solution. At higher levels the differences were not substantial. Inhibition zones of tylosin were smaller and irregular or had disappeared completely, while ceftiofur, sulfadimidine, erythromycin, lincomycin, and streptomycin were not detected in spiked muscle tissue at concentrations fivefold higher than the detection limits without tissue. These results indicate that ceftiofur, sulfonamides, streptomycin and some macrolide antibiotics cannot be detected in intact meat with the plates and bacterial strains prescribed in the European Four Plate Test, a test which was initially intended as a multi-residue method for muscle tissue. Two plates of this system are not suitable for screening purposes; a third one detects tetracyclines and beta-lactam antibiotics in spiked tissue; the fourth one is sensitive for beta-lactam antibiotics and for some but not all macrolides. Samples spiked with the fluoroquinolones enrofloxacin and ciprofloxacin can be detected with an additional plate, not included in the Four Plate Test.

Animals↗
PubMed2002

Clinical prevalence and antimicrobial susceptibility of Staphylococcus aureus and Staph. intermedius in dogs.

AIMS: This study was undertaken to investigate whether the antibiotic resistance of Staphylococcus aureus and Staph. intermedius varies with the site of isolation, sex or age of dogs. METHODS AND RESULTS: A total of 867 isolates of Staph. aureus and 1339 isolates of Staph. intermedius were obtained from nose, eye, ear, reproductive extremity, urine, abscess, skin and throat isolates. Staphylococcus intermedius isolates were isolated most frequently and adult and male dogs were more common compared with juveniles and/or female dogs. Antimicrobial resistance was commonly found for penicillin G, lincomycin, tetracycline and trimethoprim-sulphamethoxazole in both Staphylococcus species. Surprisingly, we detected significant resistance to cloxacillin in male (67.1%) and female (69.4%) Staph. aureus isolates, irrespective of the anatomical site of isolation. The resistance or susceptibility of isolates of Staph. aureus from reproductive extremities and isolates of Staph. intermedius from ear, eye and abscess sites was associated with the age of the animal. CONCLUSIONS: Antimicrobial susceptibilities in Staph. aureus and Staph. intermedius often differed with regard to the site of isolation, sex and age of the animal. SIGNIFICANCE AND IMPACT OF THE STUDY: Increasing antimicrobial resistance in staphylococci in veterinary medicine complicates the empirical selection of antimicrobial agents. These complications reveal a continuously evolving, complicated multifactoral process of the site of isolation, sex and age of the animal.

Aging↗
PubMed1997

Minimum inhibitory concentrations for selected antimicrobial agents against Fusobacterium necrophorum isolated from hepatic abscesses in cattle and sheep.

Minimum inhibitory concentrations for 35 antimicrobial agents against 100 Fusobacterium necrophorum isolates from hepatic abscesses in sheep and cattle were determined. Twelve of the thirteen beta-lactam antibiotics tested inhibited growth of 100% of strains tested. Of the remaining antimicrobial agents, extensive susceptibility was found for: spiramycin, josamycin, lincomycin, tylosin, oxytetracycline, chlortetracycline, rufloxacin, metronidazole, cotrimoxazole, sulfadimethoxine, virginiamycin and fosfomycin.

Animals↗
PubMed1998

Light regulates the rate of translation elongation of chloroplast reaction center protein D1.

Intact and lysed chloroplasts isolated from the day or night phase of seedling growth exhibit a higher rate of [35S]Met incorporation into the D1 protein in the light than in darkness. In the presence of the translation initiation inhibitor lincomycin, radiolabel incorporation remains unaffected for 7.5-10 min of the in vitro translation reaction, indicating that radiolabel incorporation is regulated by translation elongation. The rate of [35S]Met incorporation into D1-protein can be increased by addition of exogenous ATP to the in vitro translation reactions; however, ATP cannot replace light, and at physiological concentrations of stromal ATP (40 microM), the rate is at least 25-fold higher in the light than in darkness. This indicates that translation elongation is arrested in darkness. Separation of translation-elongation reactions into polysome-bound and membrane-integrated D1 proteins demonstrates that the rate of translation elongation is higher in the presence of light. In the light, less time is required to transiently radiolabel a D1 translation intermediate of about 17 kDa and to chase the translation intermediate into mature D1 protein. We propose that light regulates the enzymatic activity of the translation-elongation process in chloroplasts.

Chloroplasts↗
PubMed2000

Antimicrobial susceptibility of Arcanobacterium pyogenes isolated from cattle and pigs.

The minimum inhibitory concentrations (MICs) of 18 antimicrobial agents were determined for 49 Arcanobacterium pyogenes isolates (42 bovine isolates and 7 porcine isolates). Benzylpenicillin and ampicillin were the most active antibiotics, with MIC ranges of < or = 0.0125-0.05 microgram/ml for both bovine and porcine isolates. All isolates were susceptible to penicillins and cephems. MICs for 90% of the isolates of dihydrostreptomycin, gentamicin and oxytetracycline for bovine isolates were > 100 micrograms/ml, 1.56 micrograms/ml and 25 micrograms/ml, respectively. More resistance to dihydrostreptomycin appeared among porcine isolates (85.7%) than among bovine isolates (52.4%). Resistance to gentamicin occurred in only 3 (7.1%) of the bovine isolates. Resistance to oxytetracycline also appeared more frequent among porcine isolates (85.7%) than among bovine isolates (57.1%). All bovine isolates were susceptible to erythromycin, tilmocosin and lincomycin, but two porcine isolates (28.6%) were simultaneously resistant to these antibiotics. Tiamulin was as active as tilmicosin, with an MIC for 50% of the isolates (MIC50) of 0.05 microgram/ml for both bovine and porcine isolates. The MIC50s of chloramphenicol and its derivatives florfenicol and thiamphenicol were all 1.56 micrograms/ml. The fluoroquinolones enrofloxacin and ofloxacin were not so active as penicillins and macrolides, with MIC50s of 0.78 microgram/ml and 1.56 micrograms/ml, respectively, for both bovine and porcine isolates.

Actinomycetaceae↗
PubMed2001

Antimicrobial susceptibilities of Erysipelothrix rhusiopathiae isolated from pigs with swine erysipelas in Japan, 1988-1998.

The susceptibility to 21 antimicrobial agents of 214 strains of Erysipelothrix rhusiopathiae isolated from pigs affected with swine erysipelas in Japan between 1988 and 1998 was determined. Ampicillin, cloxacillin, benzylpenicillin, ceftiofur, tylosin, enrofloxacin and danofloxacin were the most active agents [minimum inhibitory concentrations (MICs); < or = 0.025-0.78 microgram/ml], followed by cefazolin, virginiamycin, tiamulin, chloramphenicol, florphenicol and oxolinic acid (MICs; 0.1-25 micrograms/ml). Activity was poor or absent with kanamycin and sulfadimethoxine. Strains resistant to dihydrostreptomycin, erythromycin, clindamycin, lincomycin, oxytetracycline and doxycycline were detected. The susceptibilities to dihydrostreptomycin and oxytetracycline tended to decrease. Investigation of the differences in antimicrobial susceptibility of the 214 strains according to their serotypes, sources, isolation years and regions, showed that the strains resistant to dihydrostreptomycin were most frequently found in the strains of serotype 1a and in strains from septicaemic cases. Strains resistant to oxytetracycline were detected in all serotypes and all sources, and most of the strains resistant to erythromycin were detected in the strains of serotype 2. The frequency of strains resistant to dihydrostreptomycin gradually increased from 1988 to 1996, but then decreased between 1997 and 1998. The frequency of strains resistant to oxytetracycline was remained more than 38% from 1988 to 1998. It was suggested that the strains resistant to dihydrostreptomycin and oxytetracycline were distributed over almost all districts of Japan.

Animals↗
PubMed2002

Minimum inhibitory concentrations of 20 antimicrobial agents against Staphylococcus aureus isolated from bovine intramammary infections in Japan.

Minimum inhibitory concentrations (MICs) of 20 antimicrobial agents were determined against 51 isolates of Staphylococcus aureus from bovine intramammary infections. Fourteen (27.4%) isolates were resistant to benzylpenicillin, but none of the isolates was resistant to cloxacillin, nafcillin, or cephems. Among aminoglycosides, gentamicin was the most active, with an MIC50 of 0.2 microg/ml, followed by kanamycin, with an MIC50 of 0.78 microg/ml. Five isolates (9.8%) were resistant to dihydrostreptomycin, three isolates (5.9%) to kanamycin and two isolates (3.9%) to gentamicin. Resistance to erythromycin was observed in two isolates (3.9%). Tylosin was less active than erythromycin, with MIC50s of 1.56 microg/ml versus 0.39 microg/ml, but none of the isolates was resistant to this antibiotic. Oxytetracycline MICs were situated in the range of 0.39-1.56 microg/ml for 48 susceptible isolates. Although 19 (37.3%) isolates were resistant to one or more antimicrobial agents, a single resistance pattern was most frequent: benzylpenicillin (12 isolates), dihydrostreptomycin (two isolates) and kanamycin (one isolate). There were no isolates resistant to antimicrobial agents such as methicillin, lincomycin, clindamycin, chloramphenicol, florfenicol and virginiamycin, which have not been approved for use in cattle husbandry in Japan.

Animals↗
PubMed1971

Altered methylation of ribosomal RNA in an erythromycin-resistant strain of Staphylococcus aureus.

In certain strains of Staphylococcus aureus, a concentration of erythromycin between 10(-8) and 10(-7) M can induce resistance to concentrations of this drug as high as 10(-4) M. In one such strain studied, S. aureus (1206), N(6)-dimethyladenine is not normally present in 23S rRNA; however, a compound presumptively identified (on the basis of paper chromatography in three different solvents) as N(6)-dimethyladenine appears in the 23S rRNA of growing cells that have been incubated in a medium containing 10(-7) M erythromycin. It has been shown previously that the induction of the erythromycin-resistant phenotype that occurs under these conditions requires 10(-8)-10(-7) M erythromycin for maximal expression within 1 hr and that induction results in modified 50S ribosomal subunits, which are then unable to bind erythromycin or lincomycin. Methylated adenine is also found in the 16S rRNA from the strain of S. aureus studied; however, in contrast to the situation with 23S rRNA, the amount in 16S rRNA is not affected by erythromycin. These findings provide the first example of a correlation between the methylation of rRNA and altered ribosomal function.

Adenine↗
PubMed1973

Identification of chloramphenicol-binding protein in Escherichia coli ribosomes by affinity labeling.

Monoiodoamphenicol, a synthetic analogue of chloramphenicol, has been shown by competition experiments with chloramphenicol and lincomycin to bind at the same site of 70S ribosomes as chloramphenicol. At - 2 degrees it forms a 1:1 complex with 70S ribosomes having a value of K (7.5 x 10(4) M(-1)) that is one order of magnitude lower than that of chloramphenicol. At 37 degrees , monoiodoamphenicol irreversibly inhibits the protein-synthesizing activity of E. coli ribosomes. It is shown that the analogue reacted preferentially with protein L16 of E. coli 70S ribosomes, and we therefore conclude that protein L16 belongs to the chloramphenicol-binding site of E. coli ribosomes. Since the chemically reactive group of monoiodoamphenicol resembles iodoacetamide, the reaction of E. coli 70S ribosomes with monoiodoamphenicol was compared to that with iodoacetamide. Iodoacetamide did not react with protein L16, but it predominantly reacted with proteins S18 of the 30S subunit. Furthermore, monoiodoamphenicol was reacted with E. coli ribosomal subunits. Isolated 50S subunits bound monoiodoamphenicol by about one order of magnitude less than 70S ribosomes. Again, protein L16 reacted with the affinity label. Monoiodoamphenicol reacted with protein S18 in isolated 30S subunits; it also bound to 70S ribosomes of Bacillus stearothermophilus, however, it did not bind irreversibly to these 70S ribosomes.

Acetamides↗
PubMed1985

Efficient induction and selection of chloroplast-encoded antibiotic-resistant mutants in Nicotiana.

A high rate of plastome-encoded mutations was induced in Nicotiana by exposing seeds to N-nitroso-N-methylurea. Seeds then were subjected to nutritional and in vitro selection procedures for systematic isolation of plastome-dependent antibiotic-resistant plants. Multiple flowering lines resistant to streptomycin, spectinomycin, lincomycin, or chloramphenicol were obtained. A detailed analysis of the streptomycin-resistant lines is presented. Sexual hybridization, cybrid formation following protoplast fusion, and in organello protein synthesis were used to rigorously assign the mutations to the chloroplast genome. The efficient rates of mutagenesis combined with the in vitro mass-screening procedures described here should facilitate investigation of fundamental aspects of chloroplast genetics in higher plants.

Journal Article↗
PubMed1999

Genetic engineering of the unsaturation of fatty acids in membrane lipids alters the tolerance of Synechocystis to salt stress.

The role of unsaturated fatty acids in membrane lipids in the tolerance of the photosynthetic machinery to salt stress was studied by comparing the desA-/desD- mutant of Synechocystis sp. PCC 6803, which contained monounsaturated fatty acids, with the wild-type strain, which contained a full complement of polyunsaturated fatty acids. In darkness, the loss of oxygen-evolving photosystem II activity in the presence of 0.5 M NaCl or 0.5 M LiCl was much more rapid in desA-/desD- cells than in wild-type cells. Oxygen-evolving activity that had been lost during incubation with 0.5 M NaCl in darkness returned when cells were transferred to conditions that allowed photosynthesis or respiration. Recovery was much greater in wild-type than in desA-/desD- cells, and it was prevented by lincomycin. Thus, the unsaturation of fatty acids is important in the tolerance of the photosynthetic machinery to salt stress. It appears also that the activity and synthesis of the Na+/H+ antiporter system might be suppressed under high-salt conditions and that this effect can be reversed, in part, by the unsaturation of fatty acids in membrane lipids.

Cyanobacteria↗
PubMed1996

The biosynthetic pathway of the aminonucleoside antibiotic puromycin, as deduced from the molecular analysis of the pur cluster of Streptomyces alboniger.

The pur cluster which encodes the puromycin biosynthetic pathway from Streptomyces alboniger was subcloned as a 13-kilobase fragment in plasmid pIJ702 and expressed in an apparently regulated manner in the heterologous host Streptomyces lividans. The sequencing of a 9.1-kilobase DNA fragment completed the sequence of pur. This permitted identification of seven new open reading frames in the order: napH, pur7, pur10, pur6, pur4, pur5, and pur3. The latter is followed by the known pac, dmpM, and pur8 genes. Nine open reading frames are transcribed rightward as a unit in opposite direction to that of the pur8 gene which is expressed as a monocistronic transcript from the right-most end. napH encodes the known N-acetylpuromycin N-acetylhydrolase. The deduced products from other open reading frames present similarities to: NTP pyrophosphohydrolases (pur7), several oxidoreductases (pur10), the putative LmbC protein of the lincomycin biosynthetic pathway from Streptomyces lincolnensis (pur6), S-adenosylmethionine-dependent methyltransferases (pur5), a variety of presumed aminotransferases (pur4), and several monophosphatases (pur3). According to these similarities and to previous biochemical work, a puromycin biosynthetic pathway has been deduced. No cluster-associated regulatory gene was found. However, both pur10 and pur6 genes contain a TTA codon, which suggests that they are translationally controlled by the bldA gene product, a specific tRNA(Leu).

Amino Acid Sequence↗