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Applied and Environmental Microbiology, May 2007, p. 3000-3008, Vol. 73, No. 9
0099-2240/07/$08.00+0     doi:10.1128/AEM.02612-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

The dasABC Gene Cluster, Adjacent to dasR, Encodes a Novel ABC Transporter for the Uptake of N,N'-Diacetylchitobiose in Streptomyces coelicolor A3(2){triangledown}

Akihiro Saito,1* Tomonori Shinya,2 Katsushiro Miyamoto,3 Tomofumi Yokoyama,4 Hanae Kaku,2 Eiichi Minami,5 Naoto Shibuya,2 Hiroshi Tsujibo,3 Yoshiho Nagata,1 Akikazu Ando,4 Takeshi Fujii,6 and Kiyotaka Miyashita6

Department of Bioresources Chemistry, Faculty of Horticulture, Chiba University, Matsudo 648, Matsudo City, Chiba 271-8510,1 Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571,2 Department of Microbiology, Osaka University of Pharmaceutical Sciences, Takatsuki, Osaka 569-1094,3 Department of Biotechnology, Graduate School of Science and Technology, Chiba University, Matsudo 648, Matsudo City, Chiba 271-8510,4 National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602,5 National Institute of Agro-Environmental Sciences, Kannondai 3-1-1, Tsukuba, Ibaraki 305-8604, Japan6

Received 9 November 2006/ Accepted 19 February 2007

N,N'-Diacetylchitobiose [(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 (GlcNAc)2 (equilibrium dissociation constant [KD] = 3.22 x 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.

* Corresponding author. Mailing address: Department of Bioresources Chemistry, Faculty of Horticulture, Chiba University, Matsudo 648, Matsudo City, Chiba 271-8510, Japan. Phone and fax: (81-47) 308-8871. E-mail: takosaito{at}faculty.chiba-u.jp

{triangledown} Published ahead of print on 9 March 2007.

Applied and Environmental Microbiology, May 2007, p. 3000-3008, Vol. 73, No. 9
0099-2240/07/$08.00+0     doi:10.1128/AEM.02612-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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