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21.
Andreas Bechthold Jae Kyung Sohng Todd M. Smith Xin Chu Heinz G. Floss 《Molecular genetics and genomics : MGG》1995,248(5):610-620
A 50 kb region of DNA fromStreptomyces violaceoruber Tü22, containing genes encoding proteins involved in the biosynthesis of granaticin, was isolated. The DNA sequence of a 7.3 kb fragment from this region, located approximately 10 kb from the genes that encode the polyketide synthetase responsible for formation of the benzoisochromane quinone skeleton, revealed five open reading frames (ORF1-ORF5). The deduced amino acid sequence of GraE, encoded by ORF2, shows 60.8% identity (75.2% similarity) to a dTDP-glucose dehydratase (StrE) fromStreptomyces griseus. Cultures ofEscherichia coli containing plasmids with ORF2, on a 2.1 kbBamHI fragment, were able to catalyze the formation of dTDP-4-keto-6-deoxy-d-glucose from dTDP-glucose at 5 times the rate of control cultures, confirming that ORF2 encodes a dTDP-glucose dehydratase. The amino acid sequence encoded by ORF3 (GraD) is 51.4% identical (69.9% similar) to that of StrD, a dTDP-glucose synthase fromStreptomyces griseus. The amino acid sequence encoded by ORF4 shares similarities with proteins that confer resistance to tetracycline and methylenomycin, and is suggested to be involved in transporting granaticin out of the cells by an active efflux mechanism. 相似文献
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Dutko L Rebets Y Ostash B Luzhetskyy A Bechthold A Nakamura T Fedorenko V 《FEMS microbiology letters》2006,255(2):280-285
The prx gene, which is highly homologous to putative proteinases, has been identified by sequencing in the vicinity of the biosynthetic gene cluster for landomycin E (LaE) biosynthesis (lnd) in Streptomyces globisporus 1912. The S. globisporus Pro6 gene, deficient in prx, produced fivefold less LaE than the parental strain. The expression of prx in S. globisporus Pro6 restored LaE production to wild-type levels, whereas expression of the pathway-specific regulatory gene lndI did not. The introduction of additional copies of prx into the wild-type strain using a pSG5-based plasmid, pKC1139, led to a 2.7-fold increase in LaE production. These results indicate that prx is a novel regulatory gene for LaE biosynthesis. 相似文献
24.
A. Luzhetskyy M. Fedoryshyn O. Gromyko B. Ostash Y. Rebets A. Bechthold V. Fedorenko 《Russian Journal of Genetics》2006,42(5):476-481
Mobilizable shuttle plasmids containing the origin of transfer (oriT) region of plasmid F (IncFI), ColIb-P9 (IncI1), and RP4/RP1 (IncPα) were constructed to test the ability of the cognate conjugation
system to mediate gene transfer from Escherichia coli to Streptomyces. The conjugative system of the IncPα plasmids was shown to be most effective in conjugative transfer, giving peak values
of (2.7 ± 0.2) × 10−2
S. lividans TK24 exconjugants per recipient cell. To assess whether the mating-pair formation system or the DNA-processing apparatus
of the IncPα plasmids is crucial in conjugative transfer, an assay with an IncQ-based mobilizable plasmid (RSF1010) specifying
its own DNA-processing system was developed. Only the IncPα plasmid mobilized the construct to S. lividans indicating that the mating-pair formation system is primarly responsible for the promiscuous transfer of the plasmids between
E. coli and Streptomyces. Dynamic of conjugative transfer from E. coli to S. lividans was investigated and exconjugants starting from the first hour of mating were obtained.
The text was submitted by the authors in English. 相似文献
25.
Roman Makitrynskyy Bohdan Ostash Olga Tsypik Yuriy Rebets Emma Doud Timothy Meredith Andriy Luzhetskyy Andreas Bechthold Suzanne Walker Victor Fedorenko 《Open biology》2013,3(10)
Unlike the majority of actinomycete secondary metabolic pathways, the biosynthesis of peptidoglycan glycosyltransferase inhibitor moenomycin in Streptomyces ghanaensis does not involve any cluster-situated regulators (CSRs). This raises questions about the regulatory signals that initiate and sustain moenomycin production. We now show that three pleiotropic regulatory genes for Streptomyces morphogenesis and antibiotic production—bldA, adpA and absB—exert multi-layered control over moenomycin biosynthesis in native and heterologous producers. The bldA gene for tRNALeuUAA is required for the translation of rare UUA codons within two key moenomycin biosynthetic genes (moe), moeO5 and moeE5. It also indirectly influences moenomycin production by controlling the translation of the UUA-containing adpA and, probably, other as-yet-unknown repressor gene(s). AdpA binds key moe promoters and activates them. Furthermore, AdpA interacts with the bldA promoter, thus impacting translation of bldA-dependent mRNAs—that of adpA and several moe genes. Both adpA expression and moenomycin production are increased in an absB-deficient background, most probably because AbsB normally limits adpA mRNA abundance through ribonucleolytic cleavage. Our work highlights an underappreciated strategy for secondary metabolism regulation, in which the interaction between structural genes and pleiotropic regulators is not mediated by CSRs. This strategy might be relevant for a growing number of CSR-free gene clusters unearthed during actinomycete genome mining. 相似文献
26.
Iryna Ostash Bohdan Ostash riy Luzhetskyy reas Bechthold Suzanne Walker & Victor Fedorenko 《FEMS microbiology letters》2008,285(2):195-202
In Streptomyces cyanogenus S136 gene cluster for biosynthesis of polyglycosylated angucycline landomycin A (LaA), a divergently oriented gene pair for a TetR-family regulator ( lanK ) and an efflux protein ( lanJ ) is located, whose functions remained obscure. Overexpression and disruption studies showed that lanK and lanJ genes control LaA resistance. Also, a constitutive lanK overexpression led to predominant accumulation of LaA precursors bearing shorter glycoside chains. These data as well as the results of in vitro and in vivo assays of LanK activity are consistent with the idea that LanK represses lanJ and some downstream genes involved in conversion of landomycin D (a disaccharide LaA precursor) into LaA. LaA and some of its precursors accumulate in the producing cell and relieve repression by LanK, thus amplifying the biosynthesis and export of landomycins with long glycoside chains. Therefore, the main biological role of LanK appears to be the inhibition of premature extrusion of early LaA precursors from the cells, which in turn creates the optimal conditions for accumulation of LaA as the major landomycin in S. cyanogenus S136. 相似文献
27.
The elevated relative biological effectiveness (RBE) of heavy ions like carbon is the main reason for their use in radiotherapy
and is due to the microscopic distribution of dose inside each particle track. High local doses produce lesions that are expected
to have a diminished possibility of repair. Thus, RBE depends on track structure and on the biological repair capacity of
the tissue that is affected by the irradiation. For tumor treatment planning with heavy ions, the beam quality and the tissue
sensitivity have to be taken into account. Using the dependence of radial dose distribution on particle energy and atomic
number on the physical side and x-ray dose response for the repair capacity on the biological side, the response to particle
irradiation can be calculated in the local effect model (LEM) and used for treatment planning. This article traces the route
from electron emission as the basis of track structure to the RBE calculation and the application in treatment planning.
Received: 21 April 1999 / Accepted in revised form: 1 September 1999 相似文献
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Lucia Westrich Silvie Domann Bettina Faust David Bedford David A Hopwood reas Bechthold 《FEMS microbiology letters》1999,170(2):381-387
From a cosmid library of Streptomyces cyanogenus S136, DNA fragments encompassing approximately 35 kb of the presumed landomycin biosynthetic gene cluster were identified and sequenced, revealing 32 open reading frames most of which could be assigned through data base comparison. 相似文献
30.
Liliya Horbal Yuriy Rebets Maria Rabyk Andriy Luzhetskyy Bogdan Ostash Elisabeth Welle Tatsunosuke Nakamura Victor Fedorenko Andreas Bechthold 《Applied microbiology and biotechnology》2010,85(4):1069-1079
Analysis of the α-lipomycin biosynthesis gene cluster of Streptomyces aureofaciens Tü117 led to the identification of five putative regulatory genes, which are congregated into a subcluster. Analysis of the
lipReg1–4 and lipX1 showed that they encode components of two-component signal transduction systems (LipReg1 and LipReg2), multiple antibiotics
resistance-type regulator (LipReg3), large ATP-binding regulators of the LuxR family-type regulator (LipReg4), and small ribonuclease
(LipRegX1), respectively. A combination of targeted gene disruptions, complementation experiments, lipomycin production studies,
and gene expression analysis via RT-PCR suggests that all regulatory lip genes are involved in α-lipomycin production. On the basis of the obtained data, we propose that LipReg2 controls the activity
of LipReg1, which in its turn govern the expression of the α-lipomycin pathway-specific regulatory gene lipReg4. The ribonuclease gene lipX1 and the transporter regulator lipReg3 appear to work independently of genes lipReg1, lipReg2, and lipReg4. 相似文献