Giant linear plasmids in Streptomyces which code for antibiotic biosynthesis genes

Kinashi, Haruyasu; Shimaji, Miyuki; Sakai, Akira

doi:10.1038/328454a0

Cited by 191 publications

(130 citation statements)

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“…Though megaplasmids are common within this genus of actinomycetes, this type of extrachromosomal replicon is not exclusive to rhodococci. Large plasmids (>50 kb) have been found in Gram-negative bacteria (Argandona et al, 2003;Pedraza and Diaz Ricci, 2002) and other types of Gram-positive bacteria (Kinashi et al, 1987;Scholle et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

TraA is required for megaplasmid conjugation in Rhodococcus erythropolis AN12

Yang

Lessard

Sengupta

et al. 2007

Plasmid

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Pulsed-Weld gel electrophoresis (PFGE) revealed three previously uncharacterized megaplasmids in the genome of Rhodococcus erythropolis AN12. These megaplasmids, pREA400, pREA250, and pREA100, are approximately 400, 250, and 100 kb, respectively, based on their migration in pulsed-Weld gels. Genetic screening of an AN12 transposon insertion library showed that two megaplasmids, pREA400, and pREA250, are conjugative. Mobilization frequencies of these AN12 megaplasmids to recipient R. erythropolis SQ1 were determined to be approximately 7 £ 10 ¡4 and 5 £ 10 ¡4 events per recipient cell, respectively. It is known for other bacterial systems that a relaxase encoded by the traA gene is required to initiate DNA transfer during plasmid conjugation. Sequences adjacent to the transposon insertion in megaplasmid pREA400 revealed a putative traA-like open reading frame. A targeted gene disruption method was developed to generate a traA mutation in AN12, which allowed us to address the role of the traA gene product for Rhodococcus megaplasmid conjugation. We found that the AN12 traA mutant is no longer capable of transferring the pREA400 megaplasmid to SQ1. Furthermore, we conWrmed that the conjugation defect was speciWcally due to the disruption of the traA gene, as pREA400 megaplasmid conjugation defect is restored with a complementing copy of the traA gene.

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Section: Introductionmentioning

confidence: 99%

TraA is required for megaplasmid conjugation in Rhodococcus erythropolis AN12

Yang

Lessard

Sengupta

et al. 2007

Plasmid

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“…The application of pulsed-field gel electrophoresis (PFGE) to Streptomyces DNA changed this situation and revealed that SCP1 is a giant linear plasmid of about 350 kb. 10 At the same time, many linear plasmids were detected in other antibiotic-producing strains, including S. rochei and S. lasaliensis (Table 1). 4 Furthermore, subsequent physical analysis revealed that Streptomyces chromosomes themselves are linear.…”

Section: Scp1 In Streptomyces Coelicolor Isolation and Characterizatimentioning

confidence: 97%

Giant linear plasmids in Streptomyces: a treasure trove of antibiotic biosynthetic clusters

Kinashi

2010

J Antibiot

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Many giant linear plasmids have been isolated from Streptomyces by using pulsed-field gel electrophoresis and some of them were found to carry an antibiotic biosynthetic cluster(s); SCP1 carries biosynthetic genes for methylenomycin, pSLA2-L for lankacidin and lankamycin, and pKSL for lasalocid and echinomycin. Accumulated data suggest that giant linear plasmids have played critical roles in genome evolution and horizontal transfer of secondary metabolism. In this review, I summarize typical examples of giant linear plasmids whose involvement in antibiotic production has been studied in some detail, emphasizing their finding processes and interaction with the host chromosomes. A hypothesis on horizontal transfer of secondary metabolism involving giant linear plasmids is proposed at the end.

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“…3C, compound 7 Another compound, LM-KA26B (compound 8), showed a parent ion at m/z 801, suggesting that it is dideoxylankamycin. In the 13 C NMR spectra of LM-KA26B and LM, the C-8 Possible order of two hydroxylation steps. The structures of the metabolites produced by two P450 hydroxylase-deficient mutants revealed that LkmF is a C-8 hydroxylase and LkmK is a C-15 hydroxylase.…”

Section: Metabolites Of the Orf37 (Lkmk) Disruptantmentioning

confidence: 99%

“…It was revealed that pSLA2-L contains two type I PKS gene clusters for LM (lkm) and lankacidin (lkc), a cryptic type II polyketide gene cluster (roc), and a carotenoid biosynthetic gene cluster (crt). This finding is interesting because only a few cases are known where the antibiotic biosynthetic gene cluster is located on a linear plasmid (3,4,6,13).…”

mentioning

confidence: 99%

Analysis of the Loading and Hydroxylation Steps in Lankamycin Biosynthesis in Streptomyces rochei

Arakawa

Kodama

Tatsuno

et al. 2006

Antimicrob Agents Chemother

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The biosynthetic gene cluster of lankamycin (LM), a 14-member macrolide antibiotic, is encoded on the 210-kb linear plasmid pSLA2-L in Streptomyces rochei 7434AN4. LM contains a 3-hydroxy-2-butyl group at the C-13 position, which is different from an ethyl group in erythromycin. The following two possibilities could be considered for the origin of this starter moiety of LM biosynthesis: (i) an extra module exists in the biosynthetic gene cluster and loads an additional acetate molecule, or (ii) 3-hydroxy-2-butyrate or its equivalent is loaded and incorporated as a starter. The former possibility was eliminated by the complete sequencing of pSLA2-L, which showed no extra module. On the other hand, the latter was confirmed by incorporation of deuterium in [3-2 H]DL-isoleucine into the C-14 position of LM. The timing of hydroxylation reactions at the C-15 and C-8 positions of LM was studied by constructing disruptants of two P450 hydroxylase genes, lkmF (orf26) and lkmK (orf37). The lkmF disruptant produced 8-deoxylankamycin, while the lkmK disruptant produced both 15-deoxylankamycin and 8,15-dideoxylankamycin. These results clearly showed that LkmF is a C-8 hydroxylase and LkmK is a C-15 hydroxylase in LM biosynthesis and in addition suggested the order of hydroxylation steps; namely, hydroxylation may occur at first at C-15 by LkmK and then at C-8 by LkmF.

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Giant linear plasmids in Streptomyces which code for antibiotic biosynthesis genes

Cited by 191 publications

References 0 publications

TraA is required for megaplasmid conjugation in Rhodococcus erythropolis AN12

TraA is required for megaplasmid conjugation in Rhodococcus erythropolis AN12

Giant linear plasmids in Streptomyces: a treasure trove of antibiotic biosynthetic clusters

Analysis of the Loading and Hydroxylation Steps in Lankamycin Biosynthesis in Streptomyces rochei

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