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--- bacteria:t3e:xopi [2020/07/03 09:47]
rkoebnik
+++ bacteria:t3e:xopi [2020/07/09 10:16] (current)
rkoebnik
@@ Line 1: / Line 1: @@
 ====== XopI ======
-Author: [[https://www.researchgate.net/profile/Joana_Costa12|Joana Costa]] & Trainees from the 2<sup>nd</sup>  EuroXanth Training School\\
+Author: [[https://www.researchgate.net/profile/Joana_Costa12|Joana Costa]] & Trainees from the 2<sup>nd</sup>  EuroXanth Training School (Maria Laura Destefanis, [[https://www.researchgate.net/profile/Katarina_Gasic|Katarina Gašić]], [[https://www.researchgate.net/profile/G_Licciardello|Grazia Licciardello]], Tamara Popović)\\
 Internal reviewer: Isabel Rodrigues\\
 Expert reviewer: FIXME
 Class: XopI\\
-Family: Xop\\
+Family: XopI\\
-Prototype: (//Xanthomonas euvesicatoria// pv. //euvesicatoria// aka //Xanthomonas campestris// pv. //vescicatoria//; strain 85-10)\\
+Prototype: (//Xanthomonas euvesicatoria// pv. //euvesicatoria//, ex //Xanthomonas campestris// pv. //vesicatoria//; strain 85-10)\\
 RefSeq ID: [[https://www.ncbi.nlm.nih.gov/protein/WP_011346406.1|https://www.ncbi.nlm.nih.gov/protein/WP_011346406.1]]\\
 D structure: Unknown
@@ Line 15: / Line 15: @@
 === How discovered? ===
-Effector proteins (T3Es) can suppress the plant innate immunity and alter the plant metabolism to the pathogen’s advantage. The T3E XopI was identified in Xcv strain 85-10 due to a F-box motif based on the presence of a PIP (pathogen-inducible promoter) box in its promoter region. XopI secretion and translocation was shown during the interaction of Xcv with resistant pepper plants (Schulze //et al//., 2012). Moreover, interaction studies in yeast showed that XopI specifically interacts with one out of 21 //Arabidopsis thaliana// Skp1-like proteins (ASK), suggesting that upon infection, XopI integrates into particular Skp1-Cullin-F-box protein (SCF) which target proteins for ubiquitination (Salomon //et al//., 2011). A yeast-two- hybrid screen with XopI as bait identified five proteins, that presumably are involved in the regulation of stomatal movement. Silencing of two of these potential interactors confirmed that they mediate stomatal closure after PAMP treatment in //Nicotiana benthamiana//. In tomato plants, virulence of xopI knockout strains is dramatically reduced. The stomatal aperture is as well reduced, suggesting that XopI is essential for Xcv entry into the host plant apoplast. Stomata assays with stable xopI transgenic //N. benthamiana// lines showed, that XopI suppresses tomatal closure induced by different treatments, suggesting that XopI maybe affects different pathways of stomatal immunity (Nagel and Bonas, 2018).
+Effector proteins (T3Es) can suppress the plant innate immunity and alter the plant metabolism to the pathogen’s advantage. The T3E XopI was identified in //Xcv// strain 85-10 due to a F-box motif based on the presence of a PIP (pathogen-inducible promoter) box in its promoter region. XopI secretion and translocation was shown during the interaction of //Xcv// with resistant pepper plants (Schulze //et al//., 2012). Moreover, interaction studies in yeast showed that XopI specifically interacts with one out of 21 //Arabidopsis thaliana// Skp1-like proteins (ASK), suggesting that upon infection, XopI integrates into particular Skp1-Cullin-F-box protein (SCF) which target proteins for ubiquitination (Salomon //et al//., 2011). A yeast-two-hybrid screen with XopI as bait identified five proteins, that presumably are involved in the regulation of stomatal movement. Silencing of two of these potential interactors confirmed that they mediate stomatal closure after PAMP treatment in //Nicotiana benthamiana//. In tomato plants, virulence of xopI knockout strains is dramatically reduced. The stomatal aperture is as well reduced, suggesting that XopI is essential for //Xcv// entry into the host plant apoplast. Stomata assays with stable xopI transgenic //N. benthamiana// lines showed, that XopI suppresses tomatal closure induced by different treatments, suggesting that XopI maybe affects different pathways of stomatal immunity (Nagel & Bonas, 2018).
 === (Experimental) evidence for being a T3E ===
-The transcripts of XopI were amplified from Xcv derivative 85* strain, which expresses a constitutively active HrpG point mutant resulting in constitutive expression of the T3S system, suggesting co‐expression with T3S genes (Schulze //et al//., 2012). To investigate whether //xopI// was indeed T3SS dependently secreted and translocated into the plant cell, a translational fusion with the reporter protein AvrBs3D2, a derivative of the TAL effector AvrBs3 which lacks a T3S and translocation signal, was performed. Fusion of a functional T3S signal to AvrBs3D2 enables its translocation and thus the induction of the HR in pepper cultivar ECW-30R plants that harbor the corresponding resistance gene Bs3. When the bacteria were incubated in T3S medium, XopI1–140-AvrBs3D2 was detected in the culture supernatant of strain 85*, but not of 85*DhrcV, by an AvrBs3-specific antibody. These results demonstrate that the XopI effector contained functional T3S signals in the N-terminal regions (Schulze //et al//., 2012) To test for T3SS dependent translocation, Xcv strains 85* and 85*DhrcV expressing avrBs3D2 or //xopI// fusion were inoculated into leaves of AvrBs3-responsive pepper plants (ECW-30R) and the nearisogenic susceptible pepper line ECW, which lacks the Bs3 resistance gene. Derivatives of strain 85* expressing XopI1–140-AvrBs3D2 induced the HR in ECW-30R, but not in ECW. No HR induction was observed in plants infected with derivatives of strain 85*DhrcV. Taken together, these findings confirm the T3SS secretion and translocation of XopI, and thus their nature as T3Es.
+The transcripts of XopI were amplified from //Xcv// derivative 85* strain, which expresses a constitutively active HrpG point mutant resulting in constitutive expression of the T3S system, suggesting co‐expression with T3S genes (Schulze //et al//., 2012). To investigate whether //xopI// was indeed T3SS dependently secreted and translocated into the plant cell, a translational fusion with the reporter protein AvrBs3Δ2, a derivative of the TAL effector AvrBs3 which lacks a T3S and translocation signal, was performed. Fusion of a functional T3S signal to AvrBs3Δ2 enables its translocation and thus the induction of the HR in pepper cultivar ECW-30R plants that harbor the corresponding resistance gene //Bs3//. When the bacteria were incubated in T3S medium, XopI<sub>1–140</sub>-AvrBs3Δ2 was detected in the culture supernatant of strain 85*, but not of 85*Δ//hrcV//, by an AvrBs3-specific antibody. These results demonstrate that the XopI effector contained functional T3S signals in the N-terminal regions (Schulze //et al//., 2012) To test for T3SS-dependent translocation, //Xcv// strains 85* and 85*Δ//hrcV// expressing avrBs3Δ2 or //xopI// fusion were inoculated into leaves of AvrBs3-responsive pepper plants (ECW-30R) and the near-isogenic susceptible pepper line ECW, which lacks the //Bs3// resistance gene. Derivatives of strain 85* expressing XopI<sub>1–140</sub>-AvrBs3Δ2 induced the HR in ECW-30R, but not in ECW. No HR induction was observed in plants infected with derivatives of strain 85*Δ//hrcV//. Taken together, these findings confirm the T3SS secretion and translocation of XopI, and thus their nature as T3Es.
+Translocation class; classification based on HpaB dependence (Büttner //et al.//, 2006).
 === Regulation ===
-XopI is presumably controlled by both HrpG and HrpX. The HrpX-dependent induction of //xopR// has been described previously (Koebnik //et al.//, 2006). HrpG‐ and HrpX‐dependent co‐regulation with the T3S system (+, co‐regulation; −, constitutive expression). Translocation class; classification based on HpaB dependence (Büttner //et al.//, 2006).
+XopI is presumably controlled by both HrpG and HrpX. The HrpX-dependent induction of //xopR// has been described previously (Koebnik //et al.//, 2006). HrpG‐ and HrpX‐dependent co‐regulation with the T3S system.
 === Phenotypes ===
@@ Line 57: / Line 59: @@
 Üstün S, Börnke F (2014). Interactions of //Xanthomonas// type-III effector proteins with the plant ubiquitin and ubiquitin-like pathways Front. Plant Sci. 5: 736. DOI: [[https://doi.org/10.3389/fpls.2014.00736|10.3389/fpls.2014.00736]]
+===== Further reading =====
+Thieme F (2006). Genombasierte Identifizierung neuer potentieller Virulenzfaktoren von //Xanthomonas// //campestris// pv. //vesicatoria//. Doctoral Thesis, Martin-Luther-Universität Halle-Wittenberg, Germany. PDF: [[http://sundoc.bibliothek.uni-halle.de/diss-online/06/06H103/prom.pdf|http://sundoc.bibliothek.uni-halle.de/diss-online/06/06H103/prom.pdf]]

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