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--- bacteria:t3e:xopf [2020/07/03 09:08] – rkoebnik
+++ bacteria:t3e:xopf [2025/02/13 11:31] (current) – jfpothier
@@ Line 1: / Line 1: @@
-====== XopF ======
+====== The Type III Effector XopF from //Xanthomonas// ======
 Author: [[https://www.researchgate.net/profile/Leonor_Martins|Leonor Martins]]\\
 Internal reviewer: [[https://www.researchgate.net/profile/Jaime_Cubero|Jaime Cubero]]\\
-Expert reviewer: FIXME
+Expert reviewer: [[https://www.researchgate.net/profile/Kalyan_Mondal|Kalyan K Mondal]]
 Class: XopF\\
-Family: XopF1, XopF2, XopF3\\
+Families: XopF1, XopF2, XopF3\\
-Prototype: XopF (//Xanthomonas euvesicatoria// aka //Xanthomonas campestris pv. vesicatoria//; strain 85-10)\\
+Prototype: XCV0414 (XopF1) (//Xanthomonas euvesicatoria// pv. //euvesicatoria//, ex //Xanthomonas campestris// pv. //vesicatoria//; strain 85-10)\\
-RefSeq ID: XopF1 [[https://www.ncbi.nlm.nih.gov/protein/WP_011346095.1|WP_011346095.1]] (670 aa), XopF2 [[https://www.ncbi.nlm.nih.gov/protein/56121735|AAV74205.1]] (667 aa)\\
+GenBank ID (XopF1): [[https://www.ncbi.nlm.nih.gov/protein/CAJ22045.1|CAJ22045.1]] (670 aa)\\
+GenBank ID (XopF2): [[https://www.ncbi.nlm.nih.gov/protein/AAV74205.1|AAV74205.1]] (667 aa)\\
+GenBank ID (XopF3): [[https://www.ncbi.nlm.nih.gov/protein/ELQ07409.1|ELQ07409.1]] (687 aa)\\
+RefSeq ID: XopF1 [[https://www.ncbi.nlm.nih.gov/protein/WP_011346095.1|WP_011346095.1]] (670 aa), XopF2 [[https://www.ncbi.nlm.nih.gov/protein/WP_011348008.1|WP_011348008.1]] (667 aa), XopF3 [[https://www.ncbi.nlm.nih.gov/protein/WP_039005675.1|WP_039005675.1]] (687 aa)\\
+Synonym: Hpa4\\
 D structure: Unknown
 ===== Biological function =====
-How discovered?
+=== How discovered? ===
-XopF1 and XopF2 were identified by a genetic screen using AvrBs2 as a reporter protein (Roden //et al//., 2004).
+XopF1 and XopF2 were identified in a genetic screen, using a Tn//5//-based transposon construct harboring the coding sequence for the HR-inducing domain of AvrBs2, but devoid of the effectors' T3SS signal, that was randomly inserted into the genome of //X. campestris// pv. //vesicatoria// (//Xcv//) strain 85-10. The XopF1::AvrBs2 and XopF2::AvrBs2 fusion proteins triggered a //Bs2//-dependent hypersensitive response (HR) in pepper leaves (Roden //et al//., 2004).
 === (Experimental) evidence for being a T3E ===
-//Xcv// XopF1::AvrBs2 proteins triggered avrBs2-dependent hypersensitive response (HR) during infection in Bs2-resistant pepper leaves. Fragments of the //xopF1// gene are located within the //hrp// cluster of many //Xanthomonas// spp., although a complete ORF is present only in the //Xcv// and //Xanthomonas oryzae// pv. //oryzae// (//Xoo//) //hrp// clusters (Roden //et al//., 2004). XopF1 belongs to the class A effectors (Büttner //et al//., 2006). XopF2 is 59% identical and 68% similar to XopF1 when analysed with the pairwise BLAST algorithm. //xopF2// appears to be co-transcribed with ORF1. ORF1 analysis revealed characteristics shared by type III chaperones, and is suggested to encode an Xcv chaperone (Roden //et al//., 2004).
+Type III-dependent secretion of XopF1 and XopF2 was confirmed using a calmodulin-dependent adenylate cyclase reporter assay, with a Δ//hrpF// mutant strain serving as negative control (Roden //et al.//, 2004, Mondal //et al.//, 2016).
+Fragments of the //xopF1// gene are located within the //hrp// cluster of many //Xanthomonas// spp., although a complete ORF is present only in the //Xcv// and //Xanthomonas oryzae// pv. //oryzae// (//Xoo//) //hrp// clusters (Roden //et al//., 2004).
+XopF1 belongs to the class A effectors (Büttner //et al//., 2006). XopF2 is 59% identical and 68% similar to XopF1 when analysed with the pairwise BLAST algorithm. //xopF2// appears to be co-transcribed with ORF1. ORF1 analysis revealed characteristics shared by type III chaperones, and is suggested to encode an //Xcv// chaperone (Roden //et al//., 2004).
 === Regulation ===
 RT-PCR analysis revealed //xopF1// is regulated by //hrpG// and //hrpX// and that //xopF1//, //hpaD//, //hpaI// belong to the same operon. Upstream there is a PIP box which provides binding site for HrpX (Büttner //et al//., 2007).
-qRT-PCR revealed that transcript levels of 15 out of 18 tested non-TAL effector genes (as well as the regulatory genes //hrpG// and //hrpX//), including //xopF//, were significantly reduced in the //Xanthomonas oryzae// pv. //oryzae// Δ//xrvC// mutant compared with those in the wild-type strain PXO99<sup>A</sup>  (Liu //et al.//, 2016).
+qRT-PCR revealed that transcript levels of 15 out of 18 tested non-TAL effector genes (as well as the regulatory genes //hrpG// and //hrpX//), including //xopF//, were significantly reduced in the //Xanthomonas oryzae// pv. //oryzae// Δ//xrvC// mutant compared with those in the wild-type strain PXO99<sup>A</sup> (Liu //et al.//, 2016).
 === Phenotypes ===
-To study the possible virulence function of the putative //xopF1// operon encoding HpaD, HpaI, and XopF1 these three genes were deleted from the genome of //X. campestris// pv. //vesicatoria// 85-10. The resultant mutant strain 85-10Δ//EF// displayed a wild-type phenotype when infiltrated into susceptible and resistant plants. To investigate a possible functional redundancy due to homologous genes, //xopF2// and the flanking ORF //XCV2943// were also deleted in strain 85-10Δ//EF//. Since the resulting multiple mutant strain 85-10Δ//EF//Δ//xopF2// also behaved like the wild type in infection tests//, xopF1// and //xopF2// regions did not seem to play an obvious role in the bacterial interaction with the host plant. (Büttner //et al//., 2007). Later, //Xoo// XopF1 was proven to contribute to virulence in rice, as infection with //xopF1// mutant has shown a reduced lesion size comparing to wild type (Mondal //et al//., 2016). Additionally, XopF1 and XopF2 of //X. euvesicatoria// and //Xoo// seem to have a role in PTI suppression //in planta// namely by inhibiting callose deposition and by suppressing the induction of PTI marker genes, overall contributing to development of symptoms (Mondal //et al//., 2016; Popov //et al//., 2016). //Xoo// XopF1 triggers HR in non-host plants (Li //et al//., 2016).
+  * Roden et al. did not find significant growth defects of a //Xcv// Δ//xopF1// or Δ//xopF2// mutant in susceptible pepper and tomato leaves (Roden //et al.//, 2004)
+  * To study the possible virulence function of the putative //xopF1// operon encoding HpaD, HpaI, and XopF1 these three genes were deleted from the genome of //Xcv// 85-10. The resultant mutant strain 85-10Δ//EF// displayed a wild-type phenotype when infiltrated into susceptible and resistant plants. To investigate a possible functional redundancy due to homologous genes, //xopF2// and the flanking ORF //XCV2943// were also deleted in strain 85-10Δ//EF//. Since the resulting multiple mutant strain 85-10Δ//EF//Δ//xopF2// also behaved like the wild type in infection tests//, xopF1// and //xopF2// regions did not seem to play an obvious role in the bacterial interaction with the host plant (Büttner //et al//., 2007).
+  * Later, //Xoo// XopF1 was proven to contribute to virulence in rice, as infection with //xopF1// mutant has shown a reduced lesion size comparing to wild type (Mondal //et al//., 2016).
+  * Additionally, XopF1 and XopF2 of //X. euvesicatoria// and //Xoo// seem to have a role in PTI suppression //in planta//, namely by inhibiting callose deposition and by suppressing the induction of PTI marker genes, overall contributing to development of symptoms (Mondal //et al//., 2016; Popov //et al//., 2016).
+  * //Xoo// XopF1 triggered an HR in non-host plants (Li //et al//., 2016).
 === Localization ===
 XopF2 localizes in the Golgi apparatus, while XopF1 has been found in cytoplasm (Popov //et al//., 2016) and plasma membrane (Mondal //et al//., 2016). XopF1 is encoded within //hrp// region, between //hpaB// and //hrpF//, while XopF2 is encoded elsewhere in the bacterial chromosome (Roden //et al//., 2004; Büttner //et al//., 2007).
 === Enzymatic function ===
@@ Line 36: / Line 49: @@
 === Interaction partners ===
-XopF1 secretion and translocation is T3S-dependent, HpaH, HpaC and T3S chaperone HpaB are required for efficient secretion XopF1 (Büttner //et al//., 2006, 2007).
+XopF1 secretion and translocation is T3SS-dependent; HpaH, HpaC and T3S chaperone HpaB are required for efficient secretion XopF1 (Büttner //et al//., 2006, 2007).
 ===== Conservation =====
 === In xanthomonads ===
-Yes (//e.g.//, //X. oryzae//, //X. euvesicatoria//, //X. translucens//, //X. bromi//, //X. citri, X. arboricola, X. campestris pv. musacearum//).
+Yes (//e.g.//, //X. arboricola, X. bromi//, //X. citri, X. oryzae// pv. //oryzae//, //X. euvesicatoria//, //X. translucens//, //X. vasicola//). Since the G+C content of the //xopF1// gene is similar to that of the //Xcv// //hrp// gene cluster, it may be a member of a “core” group of //Xanthomonas// spp. effectors (Roden //et al.//, 2004).
 === In other plant pathogens/symbionts ===
@@ Line 61: / Line 76: @@
 Roden J, Belt B, Ross J, Tachibana T, Vargas J, Mudgett M (2004). A genetic screen to isolate type III effectors translocated into pepper cells during //Xanthomonas// infection. Proc. Natl. Acad. Sci. USA 101: 16624-16629. DOI: [[https://www.pnas.org/content/101/47/16624|10.1073/pnas.0407383101]]
+===== Further reading =====
+Salomon D, Dar D, Sreeramulu S, Sessa G (2011). Expression of //Xanthomonas campestris// pv. //vesicatoria// type III effectors in yeast affects cell growth and viability. Mol. Plant Microbe Interact. 24: 305-314. DOI: [[https://doi.org/10.1094/MPMI-09-10-0196|10.1094/MPMI-09-10-0196]]
+===== Acknowledgements =====
+This fact sheet is based upon work from COST Action CA16107 EuroXanth, supported by COST (European Cooperation in Science and Technology).

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