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PMID: 17351098 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

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 ·2007-05-00 ·Pages 3000-8

Saito A, Shinya T, Miyamoto K, Yokoyama T, Kaku H, Minami E, Shibuya N, Tsujibo H, Nagata Y, Ando A, Fujii T, Miyashita K

Abstract

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 [K(D)] = 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).

MeSH Terms
ATP-Binding Cassette Transporters/genetics,metabolism Amino Acid Sequence Base Sequence Chitinases/metabolism Culture Media Disaccharides/metabolism Gene Expression Regulation, Bacterial/genetics Genetic Vectors/genetics Immunoblotting Molecular Sequence Data Multigene Family/genetics Open Reading Frames/genetics Reverse Transcriptase Polymerase Chain Reaction Sequence Analysis, DNA Streptomyces coelicolor/genetics Surface Plasmon Resonance
Chemicals
ATP-Binding Cassette Transporters Culture Media Disaccharides N,N-diacetylchitobiose Chitinases
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Saito Akihiro
Department of Bioresources Chemistry, Faculty of Horticulture, Chiba University, Matsudo 648, Matsudo City, Chiba 271-8510, Japan. takosaito@faculty.chiba-u.jp
Shinya Tomonori
Miyamoto Katsushiro
Yokoyama Tomofumi
Kaku Hanae
Minami Eiichi
Shibuya Naoto
Tsujibo Hiroshi
Nagata Yoshiho
Ando Akikazu
Fujii Takeshi
Miyashita Kiyotaka
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
Published
2007-05-00
Epub
2007-00-09
Pages
3000-8
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC1892892
Subset
IM
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