Faculty Opinions recommendation of Characterization of the Sinorhizobium meliloti sinR/sinI locus and the production of novel N-acyl homoserine lactones.

Author(s):  
Beth Lazazzera
2002 ◽  
Vol 184 (20) ◽  
pp. 5686-5695 ◽  
Author(s):  
Melanie M. Marketon ◽  
Matthew R. Gronquist ◽  
Anatol Eberhard ◽  
Juan E. González

ABSTRACT Sinorhizobium meliloti is a soil bacterium which can establish a nitrogen-fixing symbiosis with the legume Medicago sativa. Recent work has identified a pair of genes, sinR and sinI, which represent a potential quorum-sensing system and are responsible for the production of N-acyl homoserine lactones (AHLs) in two S. meliloti strains, Rm1021 and Rm41. In this work, we characterize the sinRI locus and show that these genes are responsible for the synthesis of several long-chain AHLs ranging from 12 to 18 carbons in length. Four of these, 3-oxotetradecanoyl HL, 3-oxohexadecenoyl HL, hexadecenoyl HL, and octadecanoyl HL, have novel structures. This is the first report of AHLs having acyl chains longer than 14 carbons. We show that a disruption in sinI eliminates these AHLs and that a sinR disruption results in only basal levels of the AHLs. Moreover, the same sinI and sinR mutations also lead to a decrease in the number of pink nodules during nodulation assays, as well as a slight delay in the appearance of pink nodules, indicating a role for quorum sensing in symbiosis. We also show that sinI and sinR mutants are still capable of producing several short-chain AHLs, one of which was identified as octanoyl HL. We believe that these short-chain AHLs are evidence of a second quorum-sensing system in Rm1021, which we refer to here as the mel system, for “S. meliloti.”


Author(s):  
Jasnizat Bin Saidin ◽  
Mohd Effendy Abd Wahid ◽  
Gaël Le Pennec

Sponges together with associated bacteria form complex holobionts governed by various relationships established between the partners. Molecules of communication may participate in the homeostasis of these biological associations. Bacteria produce N-acyl homoserine lactones to communicate. In the present study, we characterized 102 cultivable bacteria isolated from the sponge Suberites domuncula at the genus level and described their production of AHLs during 96 h of culture in a Marine Broth medium. The presence of AHLs was monitored using the Escherichia coli pSB406 reporter strain. Sixty-seven bacteria were AHLs positive. Among them, 46 were related to the Gammaproteobacteria, eight to the Alphaproteobacteria, seven to the Firmicutes and six to the Flavobacteria. The kinetic production of AHLs was determined and compared with a reference bacterium, Pseudomonas aeruginosa PAO1 (pDA224). We established three profiles of AHLs production with no definitive pattern related to an order, a family or a genus of bacteria.


2006 ◽  
Vol 188 (6) ◽  
pp. 2173-2183 ◽  
Author(s):  
Guixia Hao ◽  
Thomas J. Burr

ABSTRACT Homologs of quorum-sensing luxR and luxI regulatory genes, avsR and avsI, were identified in Agrobacterium vitis strain F2/5. Compared to other LuxI proteins from related species, the deduced AvsI shows the greatest identity to SinI (71%) from Sinorhizobium meliloti Rm1021. AvsR possesses characteristic autoinducer binding and helix-turn-helix DNA binding domains and shares a high level of identity with SinR (38%) from Rm1021. Site-directed mutagenesis of avsR and avsI was performed, and both genes are essential for hypersensitive-like response (HR) and necrosis. Two hypothetical proteins (ORF1 and ORF2) that are positioned downstream of avsR-avsI are also essential for the phenotypes. Profiles of N-acyl-homoserine lactones (AHLs) isolated from the wild type and mutants revealed that disruption of avsI, ORF1, or ORF2 abolished the production of long-chain AHLs. Disruption of avsR reduces long-chain AHLs. Expression of a cloned avsI gene in A. tumefaciens strain NT1 resulted in synthesis of long-chain AHLs. The necrosis and HR phenotypes of the avsI and avsR mutants were fully complemented with cloned avsI. The addition of synthetic AHLs (C16:1 and 3-O-C16:1) complemented grape necrosis in the avsR, avsI, ORF1, and ORF2 mutants. It was determined by reverse transcriptase PCR that the expression level of avsI is regulated by avsR but not by aviR or avhR, two other luxR homologs which were previously shown to be associated with induction of a tobacco hypersensitive response and grape necrosis. We further verified that avsR regulates avsI by measuring the expression of an avsI::lacZ fusion construct.


Sensors ◽  
2017 ◽  
Vol 17 (4) ◽  
pp. 906 ◽  
Author(s):  
Léa Girard ◽  
Élodie Blanchet ◽  
Laurent Intertaglia ◽  
Julia Baudart ◽  
Didier Stien ◽  
...  

2004 ◽  
Vol 70 (6) ◽  
pp. 3715-3723 ◽  
Author(s):  
Inmaculada Llamas ◽  
Neela Keshavan ◽  
Juan E. Gonz�lez

ABSTRACT Population-density-dependent gene expression in gram-negative bacteria involves the production of signal molecules characterized as N-acyl homoserine lactones (AHLs). The synthesis of AHLs by numerous microorganisms has been identified by using biosensor strains based on the Agrobacterium tumefaciens and Chromobacterium violaceum quorum-sensing systems. The symbiotic nitrogen-fixing bacterium Sinorhizobium meliloti is rapidly becoming a model organism for the study of quorum sensing. This organism harbors at least three different quorum-sensing systems (Sin, Mel, and Tra), which play a role in its symbiotic relationship with its host plant, alfalfa. The Sin system is distinguished among them for the production of long-chain AHLs, including C18-HL, the longest AHL reported so far. In this work, we show that construction of a sinI::lacZ transcriptional fusion results in a strain that detects long-chain AHLs with exquisite sensitivity. Overexpression of the SinR regulator protein from a vector promoter increases its sensitivity without loss of specificity. We also show that the resulting indicator strain can recognize long-chain AHLs produced by unrelated bacteria such as Paracoccus denitrificans and Rhodobacter capsulatus. This S. meliloti indicator strain should serve as a tool for the specific detection of long-chain AHLs in new systems.


Gene ◽  
2019 ◽  
Vol 684 ◽  
pp. 58-69 ◽  
Author(s):  
Heng-Leong Kher ◽  
Thiba Krishnan ◽  
Vengadesh Letchumanan ◽  
Kar-Wai Hong ◽  
Kah-Yan How ◽  
...  

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