Applied Microbiology and Biotechnology

, Volume 77, Issue 3, pp 625–635

Characterization of a novel two-component regulatory system involved in the regulation of both actinorhodin and a type I polyketide in Streptomyces coelicolor

  • Yinhua Lu
  • Weihua Wang
  • Dan Shu
  • Weiwen Zhang
  • Lei Chen
  • Zhongjun Qin
  • Sheng Yang
  • Weihong Jiang
Applied Genetics and Molecular Biotechnology

DOI: 10.1007/s00253-007-1184-5

Cite this article as:
Lu, Y., Wang, W., Shu, D. et al. Appl Microbiol Biotechnol (2007) 77: 625. doi:10.1007/s00253-007-1184-5

Abstract

To seek more information on function of two-component regulatory systems (TCSs) in Streptomycescoelicolor, a dozen TCS-knockout mutants were generated, and phenotype changes were determined. One TCS (SCO5403/5404)-deleted mutant with phenotype change was obtained. Here, we report the characterization of this novel TCS, designated as RapA1/A2 (regulation of both actinorhodin and a type I polyketide), using genetic and proteomic approaches. Although growth and morphological analyses showed no difference between the knockout mutant and wild-type strain M145, a visible decrease of the production of actinorhodin (Act) was observed in rapA1/A2 mutant. The decrease can be restored by introducing rapA1/A2 genes on an integrative vector. A 2D-gel based proteomic analysis showed that knockout of rapA1/A2 resulted in reduced expression of a putative 3-oxoacyl-[acyl-carrier protein] reductase that is part of a biosynthetic cluster for a cryptic type I polyketide. Further reverse-transcriptase-polymerase chain reaction (RT-PCR) analyses confirmed that expression levels of several biosynthetic genes and the respective pathway-specific regulatory genes actII-ORF4 and kasO for these two clusters were all down-regulated in the rapA1/A2 mutant, compared to M145. Taken together, the results demonstrated that RapA1/A2 may serve as a positive regulator for biosynthesis of both Act and the uncharacterized polyketide in S. coelicolor, and the effects exerted by RapA1/A2 were dependent on the pathway-specific regulatory genes.

Keywords

Two-component regulatory system Proteomics Secondary metabolism Streptomyces coelicolor 

Supplementary material

253_2007_1184_Fig1_ESM.gif (96 kb)
Fig. S1

Alignment of the predicted amino acid sequences of RapA1 (SCO5403) with AfsQ1 and two other uncharacterized response regulators, SAV73 from S. avermitilis and ro08012 from Rhodococcus sp._RHA1. Identical and similar amino acids are shown as black and grey background, respectively. Conserved amino acids required for the formation of phosphorylation pocket are indicated by asterisks. The conserved Asp residue (D) to be phosphorylated is marked by encircled asterisk. In addition, the possible DNA-binding domain is also shown (GIF 127 kb)

253_2007_1184_Fig2_ESM.gif (217 kb)
Fig. S2

Alignment of the deduced amino acid sequences of RapA2 (SCO5404) with AfsQ2 and two other uncharacterized histidine kinases, SAV72 from S. avermitilis and ro08011 from Rhodococcus sp._RHA1. Identical and similar amino acids are shown as black and grey background, respectively. Asterisks indicate the essential residues for phosphorylation. The conserved His residue (H) is marked by encircled asterisk. Conserved motifs in the kinases, including H box, N box, and G box are also shown (GIF 290 kb)

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Yinhua Lu
    • 1
  • Weihua Wang
    • 1
  • Dan Shu
    • 1
  • Weiwen Zhang
    • 2
  • Lei Chen
    • 1
  • Zhongjun Qin
    • 1
  • Sheng Yang
    • 1
  • Weihong Jiang
    • 1
    • 3
  1. 1.Laboratory of Molecular Microbiology, Institute of Plant Physiology and EcologyShanghai Institutes for Biological Sciences, Chinese Academy of SciencesShanghaiPeople’s Republic of China
  2. 2.Center of Ecogenomics of the Biodesign InstituteArizona State UniversityTempeUSA
  3. 3.Institut Pasteur of ShanghaiChinese Academy of SciencesShanghaiPeople’s Republic of China