Both authors contributed equally to this work.
Study of the TmoS/TmoT two-component system: towards the functional characterization of the family of TodS/TodT like systems
Article first published online: 27 DEC 2011
© 2011 The Authors. Microbial Biotechnology © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd
Volume 5, Issue 4, pages 489–500, July 2012
How to Cite
Silva-Jiménez, H., García-Fontana, C., Cadirci, B. H., Ramos-González, M. I., Ramos, J. L. and Krell, T. (2012), Study of the TmoS/TmoT two-component system: towards the functional characterization of the family of TodS/TodT like systems. Microbial Biotechnology, 5: 489–500. doi: 10.1111/j.1751-7915.2011.00322.x
- Issue published online: 7 JUN 2012
- Article first published online: 27 DEC 2011
- Received 11 October, 2011; revised 17 November, 2011; accepted 18 November, 2011.
Fig. S1. Sequence alignment of TodS like sensor kinases. The following sequences were used for this alignment: TodS of Pseudomonas putida DOT-T1E; TmoS of P. mendocina KR1, NodV of Cupriavidus necator N-1; A2SDY5 of Methylibium petroleiphilum PM1; TutC of Thauera aromatica; Q479E0 of Dechloromonas aromatica RCB; StyS of P. fluorescens (StyS Pf) and StyS of Pseudomonas sp. Y2 (StyS Ps). The amino acids that form part of the intra-TodS phosphorelay (Busch et al., 2009) are highlighted by green arrows. The amino acids that are involved in effector recognition (Busch et al., 2007) are marked with red arrows. Sequences were aligned using the Clustal W multiple sequence alignment algorithm (Thompson et al. (1994) Nucleic Acids Res 22: 4673–4680) of the NPS@ server ( http://npsa-pbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=/NPSA/npsa_server.html). The GONNET protein weight matrix, a gap opening penalty of 10 and a gap extension penalty of 0.2 were used. Fully conserved amino acids are shown in red, highly conserved residues in green and weakly conserved amino acids in blue.
Fig. S2. Sequence alignment of TodT like response regulators. The following sequences were used for this alignment: TodT of Pseudomonas putida DOT-T1E; TmoT of P. mendocina KR1, StyR of Pseudomonas sp. Y2 (StyR Ps); StyR of P. fluorescens (StyR Pf); TutB of Thauera aromatica; NodW of Cupriavidus necator N-1; A2SDY4 of Methylibium petroleiphilum PM1 and Q479E1 of Dechloromonas aromatica RCB. The same alignment procedure as detailed in Fig. S1 was used. The phosphoryl group accepting aspartate in TodS is highlighted by a green arrow. The region shaded in yellow corresponds to the recognition helix of the helix–turn–helix DNA binding motif of the StyR structure (Milani et al., 2005).
Fig. S3. Alignment of the promoters PtodX and PtmoX . The three TodT binding sites and the -10 extended region are boxed (Lacal et al., 2008a,b). The transcription start is marked.
Fig. S4. Genetic environment of TodS/TodT homologues in Dechloromonas aromatica, Methylibium petroleiphilum and Cupriavidus necator. Figures were prepared using the genome data bank in ncbi (http://www.ncbi.nlm.nih.gov/genome). In D. aromatica and M. petroleiphilum genes were present on the chromosome. In the case of Cupriavidus necator genes were present on the plasmid pBB1p.
Fig. S5. Alignments of the PtodX promoter of P. putida DOT-T1E with DNA regions preceding the toluene monooxygenase clusters in Dechloromonas aromatica, Methylibium petroleiphilum and Cupriavidus necator.
Fig. S6. Plot of beta-galactosidase measurements for the promoter PtodX (A) and PtmoX (B) (taken from Fig. 5) against the association constants determined for the binding of different agonists to purified TodS (A) or TmoS (B). Values for TodS were taken from Busch et al. (2007) and values for TmoS were taken from Table 1 of this work.
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