Present address: Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095-1570, USA.
The autoinducer synthase LqsA and putative sensor kinase LqsS regulate phagocyte interactions, extracellular filaments and a genomic island of Legionella pneumophila
Article first published online: 9 FEB 2010
© 2010 Society for Applied Microbiology and Blackwell Publishing Ltd
Volume 12, Issue 5, pages 1243–1259, May 2010
How to Cite
Tiaden, A., Spirig, T., Sahr, T., Wälti, M. A., Boucke, K., Buchrieser, C. and Hilbi, H. (2010), The autoinducer synthase LqsA and putative sensor kinase LqsS regulate phagocyte interactions, extracellular filaments and a genomic island of Legionella pneumophila. Environmental Microbiology, 12: 1243–1259. doi: 10.1111/j.1462-2920.2010.02167.x
- Issue published online: 23 APR 2010
- Article first published online: 9 FEB 2010
- Received 18 June, 2009; accepted 15 Decmeber, 2009.
Fig. S1. Growth of L. pneumophila ΔlqsA or ΔlqsS mutant strains in AYE broth or within A. castellanii. A. Legionella pneumophila wild-type, ΔlqsA or ΔlqsS mutant strains were grown in AYE broth at 37°C for the time indicated. B. Acanthamoeba castellanii was infected at an moi of 1 with L. pneumophila wild-type (●), ΔlqsA (△), ΔlqsS (◊) or ΔicmT (■) mutant bacteria. Intracellular replication was determined by cfu at the time points indicated. C. Acanthamoeba castellanii amoebae were infected at an moi of 50 with wild-type, ΔlqsA, ΔlqsS, ΔlqsR or ΔicmT mutant L. pneumophila harbouring a vector constitutively expressing GFP (pNT-28). The percentage of infected amoebae was quantified by flow cytometry. The data shown are representative of two independent experiments (A and B) or means and standard deviations of duplicates and representative of at least three independent experiments (C); P-value < 0.05.
Fig. S2. Graphic representation of coordinate gene regulation by lqsS in a 133 kb genomic island of L. pneumophila strain Philadelphia-1. The 133 kb high G+C content locus of genomic plasticity is located adjacent to the tRNAThr gene lpg0972 and flanked by putative DNA-mobilizing or phage elements (green). The locus includes region I (putative pili components; black), which is separated by lvrB1-lvrA1 (lpg1004-1005) from region II, encoding an FoF1 ATP synthase (black) and multiple metal ion efflux systems (blue). Numbers above the genes indicate the fold change in expression of 67 out of 125 genes, which are upregulated in absence of lqsS. The position of the sinR family transcription factor gene (red) annotated only in the genome of L. pneumophila strain Paris but present also in strain Philadelphia-1 is indicated.
Fig. S3. Detection of α-hydroxyketone activity by a V. cholerae luciferase reporter strain. The V. cholerae luciferase reporter strain MM879 lacking the sensor kinase CqsS was transformed with pTS-7 (cqsS), pTS-3 (lqsS), pMW-1 (lqsS–cqsS hybrid 1), pMW-2 (lqsS–cqsS hybrid 2) or pTS-10 (vector control). Luminescence was determined after induction of the genes with IPTG for 6 h. The data shown are means and standard deviations of triplicates of independent transformants and representative of two separate experiments.
Table S1. Differential gene expression in the L. pneumophila ΔlqsA, ΔlqsS, the wild-type/pNT-36 and the ΔlqsS/pNT-36 strains. Genes (A) induced or (B) repressed in stationary growth phase in the ΔlqsS mutant strain compared with L. pneumophila wild-type strain JR32. Genes induced in stationary growth phase in (C) the JR32/pNT-36 strain compared with the ΔlqsA mutant strain, or (D) the ΔlqsS/pNT-36 strain compared with the ΔlqsS mutant strain.
Table S2. Oligonucleotides used in this study.
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Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.