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bacteria:t3e:xopi [2025/02/21 13:02] – [Biological function] rkoebnikbacteria:t3e:xopi [2025/02/24 16:35] (current) – [The Type III Effector XopI from //Xanthomonas//] rkoebnik
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 ====== The Type III Effector XopI from //Xanthomonas// ====== ====== The Type III Effector XopI from //Xanthomonas// ======
  
-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)\\ +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+Internal reviewer: [[https://www.researchgate.net/profile/Isabel-Rodrigues-12|Isabel Rodrigues]]
  
 Class: XopI\\ Class: XopI\\
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 === (Experimental) evidence for being a T3E === === (Experimental) evidence for being a T3E ===
  
-The transcripts of //xopI// were amplified from //Xcv// derivative 85* strain, which expresses the constitutively active HrpG*  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.+The transcripts of //xopI// were amplified from //Xcv// derivative 85* strain, which expresses the constitutively active HrpG* 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).+XopI belongs to the translocation class of T3SS-secreted proteins, based on HpaB dependence (Büttner //et al.//, 2006).
 === Regulation === === Regulation ===
  
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 === Localization === === Localization ===
  
-XopI is translocated by the 85*Δ//hpaB// strain, that belong to the class B.+XopI is translocated by the 85*Δ//hpaB// strain, thus belonging to the class B (Büttner //et al.//, 2006).
 === Enzymatic function === === Enzymatic function ===
  
-These phenotypes can be ascribed either to the virulence activity of the effectors in plant cells, or to their recognition by the plant surveillance system. As shown in [[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4883825/figure/f0001/|**Fig. 1A**]] and [[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4883825/table/t0001/|**Table 1**]], the XopE1, XopF2, XopH, **XopI**, XopM, XopQ, XopV, AvrBs1 and AvrXv4 effectors partially or fully inhibited cell death triggered by at least one of the cell death inducers.+The T3Es XopE1, XopF2, XopH, **XopI**, XopM, XopQ, XopV, AvrBs1 and AvrXv4 partially or fully inhibited cell death triggered by at least one of the cell death inducers (Teper //et al.//, 2015).
 === Interaction partners === === Interaction partners ===
  
-XopR and XopS belong to //Xcv// translocation class A, comprising T3Es whose translocation into plant cells is completely dependent on HpaB, whereas XopB, XopG, **XopI**, XopK, XopM and XopV were assigned to class B, because they are still translocated in the absence of HpaB (Büttner //et al.//, 2006). Both new class A effectors lack homology to known proteins or motifs, so that their molecular function remains elusive. By contrast, the class B effectors comprise the putative enzyme XopG, a member of the HopH family of putative zinc metalloproteases. Other effectors possess interesting features, for example XopI contains an F‐box motif typical for eukaryotic proteins playing a role in the ubiquitin‐26S proteasome system (UPS). The UPS controls protein stability in eukaryotes and appears to be a favorable target for many T3Es, for example members of the GALA family, which strongly contribute to the virulence of //R. solanacearum// and the E3 ubiquitin ligase AvrPtoB from //P. syringae.//+XopR and XopS belong to //Xcv// translocation class A, comprising T3Es whose translocation into plant cells depends on HpaB, whereas XopB, XopG, **XopI**, XopK, XopM and XopV were assigned to class B, because they are still translocated in the absence of HpaB (Büttner //et al.//, 2006). Both new class A effectors lack homology to known proteins or motifs, so that their molecular function remains elusive. By contrast, the class B effectors comprise the putative enzyme XopG, a member of the HopH family of putative zinc metalloproteases. Other effectors possess interesting features, for example XopI contains an F‐box motif typical for eukaryotic proteins playing a role in the ubiquitin‐26S proteasome system (UPS). The UPS controls protein stability in eukaryotes and appears to be a favorable target for many T3Es, for example members of the GALA family, which strongly contribute to the virulence of //R. solanacearum// and the E3 ubiquitin ligase AvrPtoB from //P. syringae.//
  
 ===== Conservation ===== ===== Conservation =====
bacteria/t3e/xopi.1740142931.txt.gz · Last modified: 2025/02/21 13:02 by rkoebnik