title: Phenotype overlap in Xylella fastidiosa is controlled by the cyclic di-GMP phosphodiesterase Eal in response to antibiotic exposure and diffusible signal factor-mediated cell-cell signaling.
abstract: Eal is an EAL domain protein in Xylella fastidiosa homologous to one involved in resistance to tobramycin in Pseudomonas aeruginosa. EAL and HD-GYP domain proteins are implicated in the hydrolysis of the secondary messenger bis-(3'-5')-cyclic dimeric GMP (cyclic di-GMP). Cell density-dependent communication mediated by a Diffusible Signal Factor (DSF) also modulates cyclic di-GMP levels in X. fastidiosa, thereby controlling the expression of virulence genes and genes involved in insect transmission. The possible linkage of Eal to both extrinsic factors such as antibiotics and intrinsic factors such as quorum sensing, and whether both affect virulence, was thus addressed. Expression of eal was induced by subinhibitory concentrations of tobramycin, and an eal deletion mutant was more susceptible to this antibiotic than the wild-type strain and exhibited phenotypes similar to those of an rpfF deletion mutant blocked in DSF production, such as hypermotility, reduced biofilm formation, and hypervirulence to grape. Consistent with that, the rpfF mutant was more susceptible than the wild-type strain to tobramycin. Therefore, we propose that cell-cell communication and antibiotic stress can apparently lead to similar modulations of cyclic di-GMP in X. fastidiosa, resulting in similar phenotypes. However, the effect of cell density is dominant compared to that of antibiotic stress, since eal is suppressed by RpfF, which may prevent inappropriate behavioral changes in response to antibiotic stress when DSF accumulates.
authors.affiliation: Department of Plant and Microbial Biology, University of California, Berkeley, Berkeley, CA, USA.
authors.forename: Alessandra A
authors.initials: AA
authors.lastname: de Souza
authors.forename: Michael
authors.initials: M
authors.lastname: Ionescu
authors.forename: Clelia
authors.initials: C
authors.lastname: Baccari
authors.forename: Aline M
authors.initials: AM
authors.lastname: da Silva
authors.forename: Steven E
authors.initials: SE
authors.lastname: Lindow
mesh_terms: D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000378:metabolism*; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D052160:Benzothiazoles; D018441:Biofilms / Q000187:drug effects / Q000254:growth & development; D002450:Cell Communication / Q000502:physiology*; D017931:DNA Primers / Q000235:genetics; D003959:Diamines; D004351:Drug Resistance / Q000235:genetics; D004926:Escherichia coli; D017353:Gene Deletion; D005816:Genetic Complementation Test; D008826:Microbial Sensitivity Tests; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D009930:Organic Chemicals; D010641:Phenotype; D011550:Pseudomonas aeruginosa / Q000201:enzymology; D011804:Quinolines; D016415:Sequence Alignment; D014031:Tobramycin / Q000494:pharmacology; D027843:Vitis / Q000382:microbiology; D044167:Xylella / Q000187:drug effects / Q000201:enzymology* / Q000472:pathogenicity* / Q000502:physiology
chemical_list: D000900:Anti-Bacterial Agents; D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D014031:Tobramycin
references.citation: Mol Plant Microbe Interact. 2005 Aug;18(8):856-68
references.pmid: 16134898
references.citation: Appl Environ Microbiol. 2009 Jan;75(2):483-92
references.pmid: 18997023
references.citation: Mol Plant Microbe Interact. 2010 Oct;23(10):1356-63
references.pmid: 20831412
references.citation: J Bacteriol. 2005 Aug;187(16):5560-7
references.pmid: 16077100
references.citation: Proc Natl Acad Sci U S A. 2006 Apr 25;103(17):6712-7
references.pmid: 16611728
references.citation: J Bacteriol. 2011 Sep;193(18):4685-98
references.pmid: 21764921
references.citation: Nature. 2003 Nov 20;426(6964):306-10
references.pmid: 14628055
references.citation: Mol Microbiol. 2005 Aug;57(3):629-39
references.pmid: 16045609
references.citation: J Bacteriol. 2010 Feb;192(4):1020-9
references.pmid: 20008070
references.citation: Curr Opin Microbiol. 2007 Feb;10(1):17-23
references.pmid: 17208514
references.citation: Appl Environ Microbiol. 2009 Jan;75(2):521-8
references.pmid: 19011051
references.citation: Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):2670-5
references.pmid: 18268318
references.citation: Microbiology (Reading). 2004 Aug;150(Pt 8):2497-2502
references.pmid: 15289546
references.citation: Genes Dev. 2004 Mar 15;18(6):715-27
references.pmid: 15075296
references.citation: FEMS Microbiol Lett. 2010 May;306(2):152-9
references.pmid: 20370836
references.citation: Proc Natl Acad Sci U S A. 2009 Dec 29;106(52):22416-20
references.pmid: 20018775
references.citation: Environ Microbiol. 2012 Aug;14(8):1817-29
references.pmid: 22040037
references.citation: Annu Rev Phytopathol. 2008;46:243-71
references.pmid: 18422428
references.citation: Plant Dis. 2002 Oct;86(10):1056-1066
references.pmid: 30818496
references.citation: Appl Environ Microbiol. 2012 Aug;78(15):5060-9
references.pmid: 22582064
references.citation: Trends Microbiol. 2005 Feb;13(2):52-6
references.pmid: 15680762
references.citation: Curr Opin Microbiol. 2004 Apr;7(2):185-91
references.pmid: 15063857
references.citation: Mol Microbiol. 2007 Jan;63(2):429-42
references.pmid: 17241199
references.citation: Nature. 2005 Aug 25;436(7054):1171-5
references.pmid: 16121184
references.citation: Mol Microbiol. 2006 Oct;62(2):537-51
references.pmid: 17020586
references.citation: Mol Plant Microbe Interact. 2012 Apr;25(4):453-62
references.pmid: 22204646
references.citation: Curr Microbiol. 2004 May;48(5):368-72
references.pmid: 15060734
references.citation: J Bacteriol. 2006 Jun;188(12):4169-82
references.pmid: 16740923
references.citation: Cell. 2010 Apr 2;141(1):107-16
references.pmid: 20303158
references.citation: Environ Microbiol. 2001 Jan;3(1):43-52
references.pmid: 11225722
references.citation: mBio. 2010 Oct 19;1(4):
references.pmid: 20978535
references.citation: Appl Environ Microbiol. 2011 Aug;77(15):5278-84
references.pmid: 21666009
references.citation: Appl Environ Microbiol. 2009 Mar;75(6):1679-87
references.pmid: 19151176
references.citation: Antimicrob Agents Chemother. 2000 Mar;44(3):640-6
references.pmid: 10681331
references.citation: PLoS One. 2011;6(11):e27943
references.pmid: 22132176
references.citation: FEMS Immunol Med Microbiol. 2010 Aug;59(3):253-68
references.pmid: 20497222
references.citation: Mol Plant Microbe Interact. 2008 Oct;21(10):1309-15
references.pmid: 18785826
references.citation: J Bacteriol. 2007 Oct;189(20):7507-10
references.pmid: 17693510
references.citation: Phytopathology. 2012 Nov;102(11):1045-53
references.pmid: 22877314
references.citation: Proc Natl Acad Sci U S A. 2010 Mar 30;107(13):5989-94
references.pmid: 20231439
references.citation: Future Microbiol. 2009 Sep;4(7):919-32
references.pmid: 19722844
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Appl Environ Microbiol. 2007 Nov;73(22):7252-8
references.pmid: 17827325
references.citation: Gene. 1995 Dec 1;166(1):175-6
references.pmid: 8529885
references.citation: Nat Rev Microbiol. 2009 Apr;7(4):263-73
references.pmid: 19287449
references.citation: Plant Dis. 2002 Jun;86(6):633-638
references.pmid: 30823237
references.citation: J Mol Biol. 1975 Nov 5;98(3):503-17
references.pmid: 1195397
references.citation: Phytopathology. 2011 Jan;101(1):77-84
references.pmid: 20822432
references.citation: Proc Natl Acad Sci U S A. 2004 Feb 10;101(6):1737-42
references.pmid: 14755059
references.citation: Annu Rev Genet. 2006;40:385-407
references.pmid: 16895465
references.citation: Mol Microbiol. 2007 Apr;64(2):281-92
references.pmid: 17378922
references.citation: J Bacteriol. 2000 Jun;182(12):3593-6
references.pmid: 10852895
references.citation: Appl Environ Microbiol. 2009 Apr;75(8):2275-83
references.pmid: 19218414
references.citation: Antimicrob Agents Chemother. 1985 Apr;27(4):619-24
references.pmid: 3923925