The <i>ramC</i> gene is required for morphogenesis in <i>Streptomyces coelicolor</i> and expressed in a cell type‐specific manner under the direct control of RamR

O’Connor, Tamara J.; Kanellis, Pamela; Nodwell, Justin R.

doi:10.1046/j.1365-2958.2002.03004.x

Cited by 73 publications

(96 citation statements)

References 39 publications

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“…To verify that the N terminus of Cyc2 without the C terminus of Cyc2 was sufficient for normal geosmin production, the C-terminally deleted gene was cloned into pSET⍀ (31) and introduced into the N-terminally deleted cyc2 mutant. Geosmin production was restored (Table 3).…”

Section: Only the Amino-terminal Sesquiterpene Synthase Domain Of Cycmentioning

confidence: 99%

PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin

Gust

Challis

Fowler

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

1,315

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Streptomycetes are high G؉C Gram-positive, antibiotic-producing, mycelial soil bacteria. The 8.7-Mb Streptomyces coelicolor genome was previously sequenced by using an ordered library of Supercos-1 clones. Here, we describe an efficient procedure for creating precise gene replacements in the cosmid clones by using PCR targeting and -Red-mediated recombination. The cloned Streptomyces genes are replaced with a cassette containing a selectable antibiotic resistance and oriTRK2 for efficient transfer to Streptomyces by RP4-mediated intergeneric conjugation. Supercos-1 does not replicate in Streptomyces, but the clones readily undergo double-crossover recombination, thus creating gene replacements. The antibiotic resistance cassettes are flanked by yeast FLP recombinase target sequences for removal of the antibiotic resistance and oriT RK2 to generate unmarked, nonpolar mutations. The technique has been used successfully by >20 researchers to mutate around 100 Streptomyces genes. As an example, we describe its application to the discovery of a gene involved in the production of geosmin, the ubiquitous odor of soil. The gene, Sco6073 (cyc2), codes for a protein with two sesquiterpene synthase domains, only one of which is required for geosmin biosynthesis, probably via a germacra-1 (10) E,5E-dien-11-ol intermediate generated by the sesquiterpene synthase from farnesyl pyrophosphate.FLP recombinase ͉ lambda Red recombinase ͉ oriT ͉ SCO5222 ͉ SCO6073

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Section: Only the Amino-terminal Sesquiterpene Synthase Domain Of Cycmentioning

confidence: 99%

PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin

Gust

Challis

Fowler

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

1,315

1,554

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“…c-Butyrolactone-receptor system in Streptomyces rochei ramCSAB operon, which contributed to the onset of aerial growth (O'Connor et al, 2002).…”

Section: Srrc (Orf74) Is a Positive Regulator In Spore Formationmentioning

confidence: 99%

γ-Butyrolactone autoregulator-receptor system involved in lankacidin and lankamycin production and morphological differentiation in Streptomyces rochei

et al. 2007

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An afsA homologue (srrX) and three c-butyrolactone receptor gene homologues (srrA, srrB and srrC) are coded on the giant linear plasmid pSLA2-L in Streptomyces rochei 7434AN4, a producer of two polyketide antibiotics, lankacidin and lankamycin. Construction of gene disruptants and their phenotypic study revealed that srrX and srrA make a c-butyrolactone receptor system in this strain. Addition of a c-butyrolactone fraction to an srrX-deficient mutant restored the production of lankacidin and lankamycin, indicating that the SrrX protein is not necessary for this event. In addition to a positive effect on antibiotic production, srrX showed a negative effect on morphological differentiation. The receptor gene srrA reversed both effects of srrX, while the second receptor gene homologue srrC had only a positive function in spore formation. Furthermore, disruption of the third homologue srrB greatly increased the production of lankacidin and lankamycin. Electron microscopic analysis showed that aerial mycelium formation stopped at a different stage in the srrA and srrC mutants. Overall, these results indicated that srrX, srrA, srrB and srrC constitute a complex regulatory system for antibiotic production and morphological differentiation in S. rochei. INTRODUCTIONThe filamentous Gram-positive soil bacteria of the genus Streptomyces are characterized by three distinct properties: linear chromosome, complex morphological differentiation and ability to produce varieties of secondary metabolites including antibiotics. Antibiotic biosynthetic genes in Streptomyces species usually form a condensed gene cluster on the chromosome. However, several giant linear plasmids encoding an antibiotic gene cluster(s) have been isolated: SCP1 in Streptomyces coelicolor A3(2) carries the biosynthetic gene cluster for methylenomycin (Bentley et al., 2004;Chater & Bruton, 1985;Kinashi et al., 1987;Redenbach et al., 1998), pPZG103 in Streptomyces rimosus has that for oxytetracycline (Gravius et al., 1994; Pandza et al., 1998) and pSLA2-L in Streptomyces rochei has those for lankacidin and lankamycin (Arakawa et al., 2005(Arakawa et al., , 2006Mochizuki et al., 2003;Suwa et al., 2000).S. rochei strain 7434AN4 contains three large linear plasmids (pSLA2-L, -M and -S) (Kinashi et al., 1994) and produces two structurally unrelated polyketide antibiotics, the 17-membered macrocyclic lankacidin and the 14-membered macrolide lankamycin (Fig. 1a). We have been studying the function of the largest linear plasmid pSLA2-L in antibiotic production, and finally determined its 210 614 bp nucleotide sequence (Mochizuki et al., 2003). It was revealed that pSLA2-L contains an unusually condensed gene organization for secondary metabolism (Fig. 1b); two type I PKS gene clusters for lankacidin (lkc, orf4-orf18) and lankamycin (lkm, orf24-orf53), a cryptic type II PKS gene cluster (roc, orf62-orf70) and a carotenoid biosynthetic gene cluster (crt, orf104-orf110). In addition, many regulatory genes were identified on pSLA2-L, including all the homologues in t...

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“…ramR is a response regulator gene within a cluster of genes needed for aerial hypha formation in both S. coelicolor (15,16) and S. griseus (amfR) (17). In S. coelicolor, its overexpression accelerates aerial mycelium formation in wild-type strains and restores aerial hypha formation in most defective mutants, including bldA and bldH mutants.…”

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confidence: 99%

ColonialDifferentiation in Streptomyces coelicolor Depends on Translation of a SpecificCodon within the adpA Gene

et al. 2003

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The ramC gene is required for morphogenesis in Streptomyces coelicolor and expressed in a cell type‐specific manner under the direct control of RamR

Cited by 73 publications

References 39 publications

PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin

PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin

γ-Butyrolactone autoregulator-receptor system involved in lankacidin and lankamycin production and morphological differentiation in Streptomyces rochei

ColonialDifferentiation in Streptomyces coelicolor Depends on Translation of a SpecificCodon within the adpA Gene

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