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VP963/DK1622

ABSTRACT: Phosphate regulation is complex in the developmental prokaryote Myxococcus xanthus, and requires at least three previously characterized two-component systems (TCS), designated PhoR1-PhoP1, PhoR2-PhoP2, and PhoR3-PhoP3. We report here the identification and characterization of a member of a fourth TCS, designated PhoP4, which shows high sequence similarity to the other three M. xanthus PhoP response regulators. PhoP4 is an orphan response regulator that was identified by yeast 2-hybrid screen as a possible interacting partner of PhoR2. phoP4 insertion mutation and in-frame deletion strains were constructed and assessed for developmental and phosphatase phenotypes, and then compared with the phenotypes previously reported for a _phoR2-phoP2 strain. The data indicate that the phoP4 mutations caused spore viability to be decreased by nearly two orders of magnitude, and reduced all three development-specific phosphatase activities by 80-90% under phosphate-limiting conditions. These phenotypes are stronger than those observed for mutations in the PhoR2-PhoP2 system, thereby suggesting a high degree of complexity vis-à-vis the interactions amongst the Pho signal transduction pathways in M. xanthus. Keywords: developmental analysis

ORGANISM(S): Myxococcus xanthus

PROVIDER: GSE3795 | GEO | 2005/12/10

SECONDARY ACCESSION(S): PRJNA93997

REPOSITORIES: GEO

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Project description:The two-component system (TCS) is a specific regulatory system in bacteria and plays an important role in sensing and adapting to the environment. In this study, we evaluated the roles of TCSs in the major cariogenic pathogen Streptococcus mutans in resistance to several types of bacteriocin. In a comprehensive analysis using individual TCS mutants, we found that two novel TCSs were associated with resistance against distinct lantibiotics nisin A (class I type A[I]) and nukacin ISK-1(class I type A[II]). One TCS, SMU.659-660 (designated as NsrRS), was related to resistance against nisin A produced by Lactococcus lactis ATCC 11454. The other TCS, SMU.1146-1145 (designated as LcrRS), was related to resistance against nukacin ISK-1 produced by Staphylococcus warneri ISK-1. NsrRS induced the expression of SMU.658 (designated as NsrX), which constitutes an operon with nsrRS, in response to nisin A. Inactivation of nsrX increased susceptibility to nisin A. Additionally, NsrX expression in S. mutans increased the binding affinity to nisin A compared to a no-expression strain. LcrRS induced the expression of SMU.1148-50 (lctFEG), which encodes an ABC transporter and is located upstream of lcrRS, in response to nukacin ISK-1. Inactivation of lctFEG significantly increased susceptibility to nukacin ISK-1. Electrophoretic mobility shift assay analysis revealed that NsrR and LcrR bound directly to regions upstream of nsrX and lctFEG, respectively. This is the first report that two distinct TCSs, NsrRS and LcrRS, are independently involved in resistance to nisin A and nukacin ISK-1 in S. mutans. Total of 14 samples were analyzed. Total RNA from each test strain and control described below were labeled with Alexa FluorÂ® 555 and Alexa FluorÂ® 647, respectively, and were cohybridized on a single array. Labeling and hybridization were performed once or twice independently. UA159 wild type as control vs. UA159 with nisinA or nukacin ISK-1, UA159 wild type with nisinA or nukacin ISK-1 vs. nsrRS or lcrRS deletion mutant in UA159 with nisinA or nukacin ISK-1.

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VP963/DK1622

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