scholarly journals Genetic Locus Encoding Functions Involved in Biosynthesis and Outer Membrane Localization of Xanthomonadin in Xanthomonas oryzae pv. oryzae

2002 ◽  
Vol 184 (13) ◽  
pp. 3539-3548 ◽  
Author(s):  
Ajay Kumar Goel ◽  
Lakshmi Rajagopal ◽  
Narayana Nagesh ◽  
Ramesh V. Sonti
Keyword(s):  
Outer Membrane ◽  
Polyketide Synthase ◽  
Genetic Locus ◽  
Open Reading Frames ◽  
Xanthomonas Oryzae ◽  
Membrane Localization ◽  
Acyl Carrier Proteins ◽  
Membrane Bound ◽  
First Time ◽  
Reading Frames

ABSTRACT Xanthomonadins are membrane-bound, brominated, aryl-polyene pigments specific to the genus Xanthomonas. We have characterized a genetic locus (pig) from Xanthomonas oryzae pv. oryzae which contains four open reading frames (ORFs) that are essential for xanthomonadin production. Three of these ORFs are homologous to acyl carrier proteins, dehydratases, and acyl transferases, suggesting a type II polyketide synthase pathway for xanthomonadin biosynthesis. The fourth ORF has no homologue in the database. For the first time, we report that a putative cytoplasmic membrane protein encoded in the pig locus is required for outer membrane localization of xanthomonadin in X. oryzae pv. oryzae. We also report the identification of a novel 145-bp palindromic Xanthomonas repetitive intergenic consensus element that is present in two places in the pig locus. We estimate that more than 100 copies of this element might be present in the genome of X. oryzae pv. oryzae and other xanthomonads.

scholarly journals Draft Genome Sequence of Saccharomonospora sp. Strain LRS4.154, a Moderately Halophilic Actinobacterium with the Biotechnologically Relevant Polyketide Synthase and Nonribosomal Peptide Synthetase Systems

2017 ◽  
Vol 5 (21) ◽  
Author(s):  
Scarlett Alonso-Carmona ◽  
Blanca Vera-Gargallo ◽  
Rafael R. de la Haba ◽  
Antonio Ventosa ◽  
Horacio Sandoval-Trujillo ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Polyketide Synthase ◽  
Draft Genome ◽  
Nonribosomal Peptide Synthetase ◽  
Open Reading Frames ◽  
Draft Genome Sequence ◽  
Nonribosomal Peptide ◽  
Moderately Halophilic ◽  
Peptide Synthetase ◽  
Reading Frames

ABSTRACT The draft genome sequence of Saccharomonospora sp. strain LRS4.154, a moderately halophilic actinobacterium, has been determined. The genome has 4,860,108 bp, a G+C content of 71.0%, and 4,525 open reading frames (ORFs). The clusters of PKS and NRPS genes, responsible for the biosynthesis of a large number of biomolecules, were identified in the genome.

scholarly journals Complete Genome Sequence of a Renamed Isolate, Trichoplusia ni Ascovirus 6b, from the United States

2018 ◽  
Vol 6 (10) ◽  
Author(s):  
Yuan-Yuan Liu ◽  
Wen-Fei Xian ◽  
Jin Xue ◽  
Yong-Lu Wei ◽  
Xiao-Wen Cheng ◽  
...  
Keyword(s):  
United States ◽  
Phylogenetic Relationship ◽  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Trichoplusia Ni ◽  
The United States ◽  
Open Reading Frames ◽  
First Time ◽  
Reading Frames

ABSTRACT The complete genome of Trichoplusia ni ascovirus 6b (TnAV-6b) was sequenced for the first time. The TnAV-6b isolate, which has its closest phylogenetic relationship with the TnAV-6a isolate, has a circular genome of 185,664 bp, with a G+C content of 46.0% and 178 predicted open reading frames.

scholarly journals Genetic Localization and Molecular Characterization of the nonS Gene Required for Macrotetrolide Biosynthesis inStreptomyces griseus DSM40695

2000 ◽  
Vol 44 (7) ◽  
pp. 1809-1817 ◽  
Author(s):  
Wyatt C. Smith ◽  
Longkuan Xiang ◽  
Ben Shen
Keyword(s):  
Polyketide Synthase ◽  
Isotope Labeling ◽  
Streptomyces Griseus ◽  
Open Reading Frames ◽  
A Cell ◽  
Cyclic Polyethers ◽  
The Difference ◽  
Reading Frames

ABSTRACT The macrotetrolides are a family of cyclic polyethers derived from tetramerization, in a stereospecific fashion, of the enantiomeric nonactic acid (NA) and its homologs. Isotope labeling experiments established that NA is of polyketide origin, and biochemical investigations demonstrated that 2-methyl-6,8-dihydroxynon-2E-enoic acid can be converted into NA by a cell-free preparation from Streptomyces lividans that expresses nonS. These results lead to the hypothesis that macrotetrolide biosynthesis involves a pair of enantiospecific polyketide pathways. In this work, a 55-kb contiguous DNA region was cloned from Streptomyces griseus DSM40695, a 6.3-kb fragment of which was sequenced to reveal five open reading frames, including the previously reported nonR andnonS genes. Inactivation of nonS in vivo completely abolished macrotetrolide production. Complementation of thenonS mutant by the expression of nonS intrans fully restored its macrotetrolide production ability, with a distribution of individual macrotetrolides similar to that for the wild-type producer. In contrast, fermentation of thenonS mutant in the presence of exogenous (±)-NA resulted in the production of nonactin, monactin, and dinactin but not in the production of trinactin and tetranactin. These results prove the direct involvement of nonS in macrotetrolide biosynthesis. The difference in macrotetrolide production between in vivo complementation of the nonS mutant by the plasmid-borne nonSgene and fermentation of the nonS mutant in the presence of exogenously added (±)-NA suggests that NonS catalyzes the formation of (−)-NA and its homologs, supporting the existence of a pair of enantiospecific polyketide pathways for macrotetrolide biosynthesis inS. griseus. The latter should provide a model that can be used to study the mechanism by which polyketide synthase controls stereochemistry during polyketide biosynthesis.

scholarly journals Membrane-Bound, 2-Keto-d-Gluconate-Yielding d-Gluconate Dehydrogenase from “Gluconobacter dioxyacetonicus” IFO 3271: Molecular Properties and Gene Disruption

2007 ◽  
Vol 73 (20) ◽  
pp. 6551-6556 ◽  
Author(s):  
Hirohide Toyama ◽  
Naoko Furuya ◽  
Ittipon Saichana ◽  
Yoshitaka Ano ◽  
Osao Adachi ◽  
...  
Keyword(s):  
Double Mutant ◽  
Culture Medium ◽  
Wild Type Strain ◽  
Reductase Activity ◽  
Pcr Primers ◽  
Open Reading Frames ◽  
Pcr Product ◽  
Mutant Strains ◽  
Membrane Bound ◽  
Reading Frames

ABSTRACT Most Gluconobacter species produce and accumulate 2-keto-d-gluconate (2KGA) and 5KGA simultaneously from d-glucose via GA in culture medium. 2KGA is produced by membrane-bound flavin adenine dinucleotide-containing GA 2-dehydrogenase (FAD-GADH). FAD-GADH was purified from “Gluconobacter dioxyacetonicus” IFO 3271, and N-terminal sequences of the three subunits were analyzed. PCR primers were designed from the N-terminal sequences, and part of the FAD-GADH genes was cloned as a PCR product. Using this PCR product, gene fragments containing whole FAD-GADH genes were obtained, and finally the nucleotide sequence of 9,696 bp was determined. The cloned sequence had three open reading frames (ORFs), gndS, gndL, and gndC, corresponding to small, large, and cytochrome c subunits of FAD-GADH, respectively. Seven other ORFs were also found, one of which showed identity to glucono-δ-lactonase, which might be involved directly in 2KGA production. Three mutant strains defective in either gndL or sldA (the gene responsible for 5KGA production) or both were constructed. Ferricyanide-reductase activity with GA in the membrane fraction of the gndL-defective strain decreased by about 60% of that of the wild-type strain, while in the sldA-defective strain, activity with GA did not decrease and activities with glycerol, d-arabitol, and d-sorbitol disappeared. Unexpectedly, the strain defective in both gndL and sldA (double mutant) still showed activity with GA. Moreover, 2KGA production was still observed in gndL and double mutant strains. 5KGA production was not observed at all in sldA and double mutant strains. Thus, it seems that “G. dioxyacetonicus” IFO 3271 has another membrane-bound enzyme that reacts with GA, producing 2KGA.

scholarly journals Cloning, Sequencing, and Functional Analysis of an Iterative Type I Polyketide Synthase Gene Cluster for Biosynthesis of the Antitumor Chlorinated Polyenone Neocarzilin in “Streptomyces carzinostaticus”

2004 ◽  
Vol 48 (9) ◽  
pp. 3468-3476 ◽  
Author(s):  
Miyuki Otsuka ◽  
Koji Ichinose ◽  
Isao Fujii ◽  
Yutaka Ebizuka
Keyword(s):  
Gene Cluster ◽  
Polyketide Synthase ◽  
Streptomyces Coelicolor ◽  
Open Reading Frames ◽  
Modular Organization ◽  
Type I ◽  
Orf3 Protein ◽  
Polyketide Synthase Gene ◽  
Polyketide Chain ◽  
Reading Frames

ABSTRACT Neocarzilins (NCZs) are antitumor chlorinated polyenones produced by “Streptomyces carzinostaticus” var. F-41. The gene cluster responsible for the biosynthesis of NCZs was cloned and characterized. DNA sequence analysis of a 33-kb region revealed a cluster of 14 open reading frames (ORFs), three of which (ORF4, ORF5, and ORF6) encode type I polyketide synthase (PKS), which consists of four modules. Unusual features of the modular organization is the lack of an obvious acyltransferase domain on modules 2 and 4 and the presence of longer interdomain regions more than 200 amino acids in length on each module. Involvement of the PKS genes in NCZ biosynthesis was demonstrated by heterologous expression of the cluster in Streptomyces coelicolor CH999, which produced the apparent NCZ biosynthetic intermediates dechloroneocarzillin A and dechloroneocarzilin B. Disruption of ORF5 resulted in a failure of NCZ production, providing further evidence that the cluster is essential for NCZ biosynthesis. Mechanistic consideration of NCZ formation indicates the iterative use of at least one module of the PKS, which subsequently releases its product by decarboxylation to generate an NCZ skeleton, possibly catalyzed by a type II thioesterase encoded by ORF7. This is a novel type I PKS system of bacterial origin for the biosynthesis of a reduced polyketide chain. Additionally, the protein encoded by ORF3, located upstream of the PKS genes, closely resembles the FADH2-dependent halogenases involved in the formation of halometabolites. The ORF3 protein could be responsible for the halogenation of NCZs, presenting a unique example of a halogenase involved in the biosynthesis of an aliphatic halometabolite.

scholarly journals Phloem Loading of Sorbitol in Apple (Malus domestica Borkh.): Cloning and Sequence Analysis of Potential H+/Sorbitol Symporters from a Mature Leaf cDNA Library

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 756B-756
Author(s):  
Edwin J. Reidel* ◽  
Brian G. Ayre ◽  
E. Robert Turgeon ◽  
Lailiang Cheng
Keyword(s):  
Cdna Library ◽  
Malus Domestica ◽  
Phloem Loading ◽  
Open Reading Frames ◽  
Sugar Transporter ◽  
Sugar Transporters ◽  
Apple Leaves ◽  
Complete Sequencing ◽  
Membrane Bound ◽  
Reading Frames

Sorbitol (d-glucitol) is the major end product of photosynthesis in apple (Malus domestica Borkh.), as well as the predominant phloem-translocated carbohydrate. The mechanism by which sorbitol is phloem-loaded for transport to heterotrophic sink tissues is unknown. We hypothesized that a plasma membrane-bound H+/sorbitol symporter mediates apoplastic phloem-loading of sorbitol. To discover genes potentially encoding sorbitol transporters, a cDNA library was constructed from mature `Gala' apple leaves. A homologous probe was synthesized via PCR with primers were designed against the cherry fruit sorbitol transporter, PcSot1, and using library lysate as template. From an initial plating of approximately 5 × 105 clones, twelve positives were identified after three rounds of hybridization screening. Following single-pass, 5' end sequencing, the clones were sorted into four contiguous sequences. One clone was chosen from each contig for complete sequencing. The four clones, provisionally named MdSOT1-4 (Malus domesitca Sorbitol Transporter), potentially encode full-length cDNAs for sorbitol transporters: Translated-BLAST searching (blastx) revealed that the open reading frames encode the complete Pfam sugar transporter domain, and the most significant alignments are with sequences encoding known- and putative polyol and sugar transporters.

scholarly journals Genetic Basis for Lipopolysaccharide O-Antigen Biosynthesis in Bordetellae

1999 ◽  
Vol 67 (8) ◽  
pp. 3763-3767 ◽  
Author(s):  
Andrew Preston ◽  
Andrew G. Allen ◽  
Joanna Cadisch ◽  
Richard Thomas ◽  
Kim Stevens ◽  
...  
Keyword(s):  
Insertion Sequence ◽  
Genetic Basis ◽  
Genetic Locus ◽  
Bordetella Bronchiseptica ◽  
Open Reading Frames ◽  
Bordetella Parapertussis ◽  
O Antigen ◽  
Surface Polysaccharide ◽  
Reading Frames

ABSTRACT Bordetella bronchiseptica and Bordetella parapertussis express a surface polysaccharide, attached to a lipopolysaccharide, which has been called O antigen. This structure is absent from Bordetella pertussis. We report the identification of a large genetic locus in B. bronchiseptica and B. parapertussis that is required for O-antigen biosynthesis. The locus is replaced by an insertion sequence in B. pertussis, explaining the lack of O-antigen biosynthesis in this species. The DNA sequence of the B. bronchiseptica locus has been determined and the presence of 21 open reading frames has been revealed. We have ascribed putative functions to many of these open reading frames based on database searches. Mutations in the locus in B. bronchiseptica andB. parapertussis prevent O-antigen biosynthesis and provide tools for the study of the role of O antigen in infections caused by these bacteria.

scholarly journals Identification and Characterization of a Gene Cluster for Synthesis of the Polyketide Antibiotic 2,4-Diacetylphloroglucinol from Pseudomonas fluorescens Q2-87

1999 ◽  
Vol 181 (10) ◽  
pp. 3155-3163 ◽  
Author(s):  
M. Gita Bangera ◽  
Linda S. Thomashow
Keyword(s):  
Genomic Dna ◽  
Plant Pathogens ◽  
Polyketide Synthase ◽  
Open Reading Frames ◽  
Fungal Plant Pathogens ◽  
Repressor Proteins ◽  
Flanking Regions ◽  
Identification And Characterization ◽  
Reading Frames

The polyketide metabolite 2,4-diacetylphloroglucinol (2,4-DAPG) is produced by many strains of fluorescent Pseudomonas spp. with biocontrol activity against soilborne fungal plant pathogens. Genes required for 2,4-DAPG synthesis by P. fluorescensQ2-87 are encoded by a 6.5-kb fragment of genomic DNA that can transfer production of 2,4-DAPG to 2,4-DAPG-nonproducing recipientPseudomonas strains. In this study the nucleotide sequence was determined for the 6.5-kb fragment and flanking regions of genomic DNA from strain Q2-87. Six open reading frames were identified, four of which (phlACBD) comprise an operon that includes a set of three genes (phlACB) conserved between eubacteria and archaebacteria and a gene (phlD) encoding a polyketide synthase with homology to chalcone and stilbene synthases from plants. The biosynthetic operon is flanked on either side by phlEand phlF, which code respectively for putative efflux and regulatory (repressor) proteins. Expression in Escherichia coli of phlA, phlC, phlB, andphlD, individually or in combination, identified a novel polyketide biosynthetic pathway in which PhlD is responsible for the production of monoacetylphloroglucinol (MAPG). PhlA, PhlC, and PhlB are necessary to convert MAPG to 2,4-DAPG, and they also may function in the synthesis of MAPG.

scholarly journals Lausannevirus Encodes a Functional Dihydrofolate Reductase Susceptible to Proguanil

2017 ◽  
Vol 61 (4) ◽  
Author(s):  
L. Mueller ◽  
P. M. Hauser ◽  
F. Gauye ◽  
G. Greub
Keyword(s):  
Dihydrofolate Reductase ◽  
Expression System ◽  
Open Reading Frames ◽  
Dna Viruses ◽  
Content Type ◽  
The Family ◽  
Nucleocytoplasmic Large Dna Viruses ◽  
First Time ◽  
Reading Frames

ABSTRACT Lausannevirus belongs to the family Marseilleviridae within the group of nucleocytoplasmic large DNA viruses (NCLDVs). These giant viruses exhibit unique features, including a large genome, ranging from 100 kb to 2.5 Mb and including from 150 to more than 2,500 genes, as well as the presence of genes coding for proteins involved in transcription and translation. The large majority of Lausannevirus open reading frames have unknown functions. Interestingly, a bifunctional dihydrofolate reductase-thymidylate synthase (DHFR-TS) is encoded in the Lausannevirus genome. The enzyme plays central roles in DNA precursor biosynthesis. DHFR is the pharmacological target of antifolates, such as trimethoprim, pyrimethamine, and proguanil. First, the functionality of Lausannevirus DHFR-TS was demonstrated by the successful complementation of a DHFR-deficient Saccharomyces cerevisiae strain with a plasmid expressing the heterologous gene. Additionally, using this heterologous expression system, we demonstrated the in vitro susceptibility of Lausannevirus DHFR-TS to proguanil and its resistance to pyrimethamine and trimethoprim. Proguanil may provide a unique and useful treatment if Lausannevirus proves to be a human pathogen. To our knowledge, this is the first time that a DHFR-TS has been described and characterized in an NCLDV.

scholarly journals Molecular and Genomic Analysis of Genes Encoding Surface-Anchored Proteins from Clostridium difficile

2001 ◽  
Vol 69 (5) ◽  
pp. 3442-3446 ◽  
Author(s):  
Tuomo Karjalainen ◽  
Anne-Judith Waligora-Dupriet ◽  
Marina Cerquetti ◽  
Patrizia Spigaglia ◽  
Andrea Maggioni ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Genetic Locus ◽  
Domain Architecture ◽  
Genomic Analysis ◽  
Open Reading Frames ◽  
Precursor Protein ◽  
Precursor Proteins ◽  
Genes Encoding ◽  
The One ◽  
Reading Frames

ABSTRACT The gene slpA, encoding the S-layer precursor protein in the virulent Clostridium difficile strains C253 and 79–685, was identified. The precursor protein carries a C-terminal highly conserved anchoring domain, similar to the one found in the Cwp66 adhesin (previously characterized in strain 79–685), an SLH domain, and a variable N-terminal domain mediating cell adherence. The genes encoding the S-layer precursor proteins and the Cwp66 adhesin are present in a genetic locus carrying 17 open reading frames, 11 of which encode a similar two-domain architecture, likely to include surface-anchored proteins.

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