The dasABC gene cluster, adjacent to dasR, encodes a novel ABC transporter for the uptake of N,N′-diacetylchitobiose in Streptomyces coelicolor A3(2)

Akihiro Saito, Tomonori Shinya, Katsushiro Miyamoto, Tomofumi Yokoyama, Hanae Kaku, Eiichi Minami, Naoto Shibuya, Hiroshi Tsujibo, Yoshiho Nagata, Akikazu Ando, Takeshi Fujii, Kiyotaka Miyashita

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

N,N′-Biacetylchitobiose [(GlcNAc)2] induces the transcription of chitinase (chi) genes in Streptomyces coelicolor A3(2). Physiological studies showed that (GlcNAc)2 addition triggered chi expression and increased the rate of (GlcNAc)2 concentration decline in culture supernatants of mycelia already cultivated with (GlcNAc)2, suggesting that (GlcNAc)2 induced the synthesis of its own uptake system. Four open reading frames (SCO0531, SCO0914, SCO2946, and SCO5232) encoding putative sugar-binding proteins of ABC transporters were found in the genome by probing the 12-bp repeat sequence required for regulation of chi transcription. SCO5232, named dasA, showed transcriptional induction by (GlcNAc)2 and N,N′,N″-triacetylchitotriose [(GlcNAc) 3]. Surface plasmon resonance analysis showed that recombinant DasA protein exhibited the highest affinity for (GlNAc)2 (equilibrium dissociation constant [KD] = 3.22 × 10-8). In the dasA-null mutant, the rate of decline of the (GlcNAc)2 concentration in the culture supernatant was about 25% of that in strain M145. The in vitro and in vivo data clearly demonstrated that dasA is involved in (GlcNAc) 2 uptake. Upstream and downstream of dasA, the transcriptional regulator gene (dasR) and two putative integral membrane protein genes (dasBC) are located in the opposite and same orientations, respectively. The expression of dasR and dasB, which seemed independent of dasA transcription, was also induced by (GlcNAc)2 and (GlcNAc)3.

Original languageEnglish
Pages (from-to)3000-3008
Number of pages9
JournalApplied and Environmental Microbiology
Volume73
Issue number9
DOIs
Publication statusPublished - May 2007
Externally publishedYes

Fingerprint

Streptomyces coelicolor
ABC transporters
ATP-Binding Cassette Transporters
Multigene Family
chitinase
multigene family
transcription (genetics)
protein
gene
surface plasmon resonance
carbohydrate binding
Chitinases
regulator genes
recombinant proteins
membrane proteins
mycelium
open reading frames
binding proteins
sugar
genes

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

The dasABC gene cluster, adjacent to dasR, encodes a novel ABC transporter for the uptake of N,N′-diacetylchitobiose in Streptomyces coelicolor A3(2). / Saito, Akihiro; Shinya, Tomonori; Miyamoto, Katsushiro; Yokoyama, Tomofumi; Kaku, Hanae; Minami, Eiichi; Shibuya, Naoto; Tsujibo, Hiroshi; Nagata, Yoshiho; Ando, Akikazu; Fujii, Takeshi; Miyashita, Kiyotaka.

In: Applied and Environmental Microbiology, Vol. 73, No. 9, 05.2007, p. 3000-3008.

Research output: Contribution to journalArticle

Saito, A, Shinya, T, Miyamoto, K, Yokoyama, T, Kaku, H, Minami, E, Shibuya, N, Tsujibo, H, Nagata, Y, Ando, A, Fujii, T & Miyashita, K 2007, 'The dasABC gene cluster, adjacent to dasR, encodes a novel ABC transporter for the uptake of N,N′-diacetylchitobiose in Streptomyces coelicolor A3(2)', Applied and Environmental Microbiology, vol. 73, no. 9, pp. 3000-3008. https://doi.org/10.1128/AEM.02612-06
Saito, Akihiro ; Shinya, Tomonori ; Miyamoto, Katsushiro ; Yokoyama, Tomofumi ; Kaku, Hanae ; Minami, Eiichi ; Shibuya, Naoto ; Tsujibo, Hiroshi ; Nagata, Yoshiho ; Ando, Akikazu ; Fujii, Takeshi ; Miyashita, Kiyotaka. / The dasABC gene cluster, adjacent to dasR, encodes a novel ABC transporter for the uptake of N,N′-diacetylchitobiose in Streptomyces coelicolor A3(2). In: Applied and Environmental Microbiology. 2007 ; Vol. 73, No. 9. pp. 3000-3008.
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abstract = "N,N′-Biacetylchitobiose [(GlcNAc)2] induces the transcription of chitinase (chi) genes in Streptomyces coelicolor A3(2). Physiological studies showed that (GlcNAc)2 addition triggered chi expression and increased the rate of (GlcNAc)2 concentration decline in culture supernatants of mycelia already cultivated with (GlcNAc)2, suggesting that (GlcNAc)2 induced the synthesis of its own uptake system. Four open reading frames (SCO0531, SCO0914, SCO2946, and SCO5232) encoding putative sugar-binding proteins of ABC transporters were found in the genome by probing the 12-bp repeat sequence required for regulation of chi transcription. SCO5232, named dasA, showed transcriptional induction by (GlcNAc)2 and N,N′,N″-triacetylchitotriose [(GlcNAc) 3]. Surface plasmon resonance analysis showed that recombinant DasA protein exhibited the highest affinity for (GlNAc)2 (equilibrium dissociation constant [KD] = 3.22 × 10-8). In the dasA-null mutant, the rate of decline of the (GlcNAc)2 concentration in the culture supernatant was about 25{\%} of that in strain M145. The in vitro and in vivo data clearly demonstrated that dasA is involved in (GlcNAc) 2 uptake. Upstream and downstream of dasA, the transcriptional regulator gene (dasR) and two putative integral membrane protein genes (dasBC) are located in the opposite and same orientations, respectively. The expression of dasR and dasB, which seemed independent of dasA transcription, was also induced by (GlcNAc)2 and (GlcNAc)3.",
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AU - Shinya, Tomonori

AU - Miyamoto, Katsushiro

AU - Yokoyama, Tomofumi

AU - Kaku, Hanae

AU - Minami, Eiichi

AU - Shibuya, Naoto

AU - Tsujibo, Hiroshi

AU - Nagata, Yoshiho

AU - Ando, Akikazu

AU - Fujii, Takeshi

AU - Miyashita, Kiyotaka

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