The pathogenesis-related 1 (PR1) proteins are members of the cross-kingdom conserved CAP superfamily (from Cysteine-rich secretory protein, Antigen 5, and PR1 proteins). PR1 mRNA expression is frequently used for biotic stress monitoring in plants; however, the molecular mechanisms of its cellular processing, localization, and function are still unknown. To analyse the localization and immunity features of Arabidopsis thaliana PR1, we employed transient expression in Nicotiana benthamiana of the tagged full-length PR1 construct, and also disrupted variants with C-terminal truncations or mutations. We found that en route from the endoplasmic reticulum, the PR1 protein transits via the multivesicular body and undergoes partial proteolytic processing, dependent on an intact C-terminal motif. Importantly, only nonmutated or processing-mimicking variants of PR1 are secreted to the apoplast. The C-terminal proteolytic cleavage releases a protein fragment that acts as a modulator of plant defence responses, including localized cell death control. However, other parts of PR1 also have immunity potential unrelated to cell death. The described modes of the PR1 contribution to immunity were found to be tissue-localized and host plant ontogenesis dependent.

Department of Biochemistry and Microbiology Faculty of Food and Biochemical Technology University of Chemistry and Technology Prague Czech Republic

Department of Experimental Plant Biology Faculty of Science Charles University Prague Czech Republic

Institute of Experimental Botany of the Czech Academy of Sciences Prague Czech Republic

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Agrios, G.N. (2005) How plants defend themselves against pathogens. In: Agrios, G.N. (Ed.), Plant Pathology. Academic Press, 207–248.

Antoniw, J.F. & Pierpoint, W.S. (1978) Purification of a tobacco leaf protein associated with resistance to virus infection. Biochemical Society Transactions, 6, 248–250. PubMed

Betsuyaku, S. , Katou, S. , Takebayashi, Y. , Sakakibara, H. , Nomura, N. & Fukuda, H. (2018) Salicylic acid and jasmonic acid pathways are activated in spatially different domains around the infection site during effector‐triggered immunity in PubMed PMC

Boudart, G. , Jamet, E. , Rossignol, M. , Lafitte, C. , Borderies, G. , Jauneau, A. et al. (2005) Cell wall proteins in apoplastic fluids of PubMed

Bourdais, G. , McLachlan, D.H. , Rickett, L.M. , Zhou, J. , Siwoszek, A. , Häweker, H. et al. (2019) The use of quantitative imaging to investigate regulators of membrane trafficking in PubMed

Bowling, S.A. , Guo, A. , Cao, H. , Gordon, A.S. , Klessig, D.F. & Dong, X. (1994) A mutation in PubMed PMC

Breen, S. , Williams, S.J. , Outram, M. , Kobe, B. & Solomon, P.S. (2017) Emerging insights into the functions of pathogenesis‐related protein 1. Trends in Plant Science, 22, 871–879. PubMed

Breen, S. , Williams, S.J. , Winterberg, B. , Kobe, B. & Solomon, P.S. (2016) Wheat PR‐1 proteins are targeted by necrotrophic pathogen effector proteins. The Plant Journal, 88, 13–25. PubMed

Camacho Henriquez, A. & Sänger, H.L. (1982) Analysis of acid‐extractable tomato leaf proteins after infection with a viroid, two viruses and a fungus and partial purification of the “pathogenesis‐related” protein p14. Archives of Virology, 74, 181–196. PubMed

Carella, P. , Merl‐Pham, J. , Wilson, D.C. , Dey, S. , Hauck, S.M. , Vlot, A.C. et al. (2016) Comparative proteomics analysis of phloem exudates collected during the induction of systemic acquired resistance. Plant Physiology, 171, 1495–1510. PubMed PMC

Cerovska, N. , Hoffmeisterova, H. , Moravec, T. , Plchova, H. , Folwarczna, J. , Synkova, H. et al. (2012) Transient expression of Human papillomavirus type 16 L2 epitope fused to N‐ and C‐terminus of coat protein of Potato virus X in plants. Journal of Biosciences, 37, 125–133. PubMed

Chen, Y.L. , Lee, C.Y. , Cheng, K.T. , Chang, W.H. , Huang, R.N. , Nam, H.G. et al. (2014) Quantitative peptidomics study reveals that a wound‐induced peptide from PR‐1 regulates immune signaling in tomato. The Plant Cell, 26, 4135–4148. PubMed PMC

Chien, P.S. , Nam, H.G. & Chen, Y.R. (2015) A salt‐regulated peptide derived from the CAP superfamily protein negatively regulates salt‐stress tolerance in PubMed PMC

Chung, K.P. , Zeng, Y. , Li, Y. , Ji, C. , Xia, Y. & Jiang, L. (2018) Signal motif‐dependent ER export of the Qc‐SNARE BET12 interacts with MEMB12 and affects PR1 trafficking in PubMed

Dixon, D.C. , Cutt, J.R. & Klessig, D.F. (1991) Differential targeting of the tobacco PR‐1 pathogenesis‐related proteins to the extracellular space and vacuoles of crystal idioblasts. The EMBO Journal, 10, 1317–1324. PubMed PMC

Du, Y. , Mpina, M.H. , Birch, P.R.J. , Bouwmeester, K. & Govers, F. (2015) PubMed PMC

Ebine, K. , Fujimoto, M. , Okatani, Y. , Nishiyama, T. , Goh, T. , Ito, E. et al. (2011) A membrane trafficking pathway regulated by the plant‐specific RAB GTPase ARA6. Nature Cell Biology, 13, 853–859. PubMed

Feige, J.N. , Sage, D. , Wahli, W. , Desvergne, B. & Gelman, L. (2005) PixFRET, an ImageJ plug‐in for FRET calculation that can accommodate variations in spectral bleed‐throughs. Microscopy Research and Technique, 68, 51–58. PubMed

Gamir, J. , Darwiche, R. , van't Hof, P. , Choudhary, V. , Stumpe, M. , Schneiter, R. et al. (2017) The sterol‐binding activity of PATHOGENESIS‐RELATED PROTEIN 1 reveals the mode of action of an antimicrobial protein. The Plant Journal, 89, 502–509. PubMed

Ge, Z. , Zhao, Y. , Liu, M.‐C. , Zhou, L.‐Z. , Wang, L. , Zhong, S. et al. (2019) LLG2/3 are co‐receptors in BUPS/ANX‐RALF signaling to regulate PubMed PMC

Gibbs, G.M. , Roelants, K. & O’Bryan, M.K. (2008) The CAP superfamily: cysteine‐rich secretory proteins, antigen 5, and pathogenesis‐related 1 proteins – roles in reproduction, cancer, and immune defense. Endocrine Reviews, 29, 865–897. PubMed

Gu, Y. & Innes, R.W. (2012) The KEEP ON GOING protein of PubMed PMC

Hála, M. , Cole, R. , Synek, L. , Drdová, E. , Pečenková, T. , Nordheim, A. et al. (2008) An exocyst complex functions in plant cell growth in PubMed PMC

Hansen, L.L. & Nielsen, M.E. (2017) Plant exosomes: using an unconventional exit to prevent pathogen entry?. Journal of Experimental Botany, 69, 59–68. PubMed

Hecker, A. , Wallmeroth, N. , Peter, S. , Blatt, M.R. , Harter, K. & Grefen, C. (2015) Binary 2in1 vectors improve in planta (co)localization and dynamic protein interaction studies. Plant Physiology, 168, 776–787. PubMed PMC

Hooft van Huijsduijnen, R.A.M. , Kauffmann, S. , Brederode, F.T. , Cornelissen, B.J.C. , Legrand, M. , Fritig, B. et al. (1987) Homology between chitinases that are induced by TMV infection of tobacco. Plant Molecular Biology, 9, 411–420. PubMed

Inada, N. , Betsuyaku, S. , Shimada, T.L. , Ebine, K. , Ito, E. , Kutsuna, N. et al. (2016) Modulation of plant RAB GTPase‐mediated membrane trafficking pathway at the interface between plants and obligate biotrophic pathogens. Plant and Cell Physiology, 57, 1854–1864. PubMed

Ito, E. , Uemura, T. , Ueda, T. & Nakano, A. (2016) Distribution of RAB5‐positive multivesicular endosomes and the trans‐Golgi network in root meristematic cells of PubMed PMC

van Loon, L.C. & van Kammen, A. (1970) Polyacrylamide disc electrophoresis of the soluble leaf proteins from PubMed

van Loon, L.C. , Rep, M. & Pieterse, C.M.J. (2006) Significance of inducible defense‐related proteins in infected plants. Annual Review of Phytopathology, 44(1), 135–162. PubMed

van Loon, L.C. & van Strien, E.A. (1999) The families of pathogenesis‐related proteins, their activities, and comparative analysis of PR‐1 type proteins. Physiological and Molecular Plant Pathology, 55, 85–97.

Jelitto‐Van Dooren, E.P.W.M. , Vidal, S. & Denecke, J. (1999) Anticipating endoplasmic reticulum stress: a novel early response before pathogenesis‐related gene induction. The Plant Cell, 11, 1935–1943. PubMed PMC

Jia, T. , Gao, C. , Cui, Y. , Wang, J. , Ding, Y. , Cai, Y. et al. (2013) ARA7(Q69L) expression in transgenic PubMed PMC

Joosten, M.H.A.J. (2012) Isolation of apoplastic fluid from leaf tissue by the vacuum infiltration‐centrifugation technique. Methods in Molecular Biology, 835, 603–610. PubMed

Joshi, V. , Joshi, N. , Vyas, A. & Jadhav, S.K. (2021) Pathogenesis‐related proteins: role in plant defense. In: Jogaiah, S. (Ed.), Biocontrol agents and secondary metabolites. Woodhead Publishing, pp. 573–590.

Jung, H.W. & Hwang, B.K. (2000) Isolation, partial sequencing, and expression of pathogenesis‐related cDNA genes from pepper leaves infected by PubMed

Kalde, M. , Nühse, T.S. , Findlay, K. & Peck, S.C. (2007) The syntaxin SYP132 contributes to plant resistance against bacteria and secretion of pathogenesis‐related protein 1. Proceedings of the National Academy of Sciences of the United States of America, 104, 11850–11855. PubMed PMC

Kama, R. , Gabriely, G. , Kanneganti, V. , Gerst, J.E. & Gilmore, R. (2018) Cdc48 and ubiquilins confer selective anterograde protein sorting and entry into the multivesicular body in yeast. Molecular Biology of the Cell, 29, 948–963. PubMed PMC

Karimi, M. , Inzé, D. & Depicker, A. (2002) GATEWAY PubMed

Klahre, U. , Becker, C. , Schmitt, A.C. & Kost, B. (2006) Nt‐RhoGDI2 regulates Rac/Rop signaling and polar cell growth in tobacco pollen tubes. The Plant Journal, 46, 1018–1031. PubMed

Kus, J.V. , Zaton, K. , Sarkar, R. & Cameron, R.K. (2002) Age‐related resistance in PubMed PMC

Lincoln, J.E. , Sanchez, J.P. , Zumstein, K. & Gilchrist, D.G. (2018) Plant and animal PR1 family members inhibit programmed cell death and suppress bacterial pathogens in plant tissues. Molecular Plant Pathology, 19, 2111–2123. PubMed PMC

Lu, S. , Faris, J.D. , Sherwood, R. & Edwards, M.C. (2013) Dimerization and protease resistance: new insight into the function of PR‐1. Journal of Plant Physiology, 170, 105–110. PubMed

Nakagawa, T. , Kurose, T. , Hino, T. , Tanaka, K. , Kawamukai, M. , Niwa, Y. et al. (2007) Development of series of Gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation. Journal of Bioscience and Bioengineering, 104, 34–41. PubMed

Nakano, R.T. , Yamada, K. , Bednarek, P. , Nishimura, M. & Hara‐Nishimura, I. (2014) ER bodies in plants of the Brassicales order: biogenesis and association with innate immunity. Frontiers in Plant Science, 5, 73. PubMed PMC

Niderman, T. , Genetet, I. , Bruyère, T. , Gees, R. , Stintzi, A. , Legrand, M. et al. (1995) Pathogenesis‐related PR‐1 proteins are antifungal: isolation and characterization of three 14‐kilodalton proteins of tomato and of a basic PR‐1 of tobacco with inhibitory activity against PubMed PMC

Niehl, A. , Amari, K. , Gereige, D. , Brandner, K. , Mély, Y. & Heinlein, M. (2012) Control of Tobacco mosaic virus movement protein fate by CELL‐DIVISION‐CYCLE protein48. Plant Physiology, 160, 2093–2108. PubMed PMC

Ortmannová, J. , Sekereš, J. , Kulich, I. , Šantrůček, J. , Dobrev, P. , Žárský, V. et al. (2021) The EXO70B2 exocyst subunit contributes to papillae and encasement formation in anti‐fungal defence in PubMed

Pečenková, T. , Pleskot, R. & Žárský, V. (2017) Subcellular localization of PubMed PMC

Pejchar, P. , Sekereš, J. , Novotný, O. , Žárský, V. & Potocký, M. (2020) Functional analysis of phospholipase Dδ family in tobacco pollen tubes. The Plant Journal, 103, 212–226. PubMed

Pruss, G.J. , Nester, E.W. & Vance, V. (2008) Infiltration with PubMed

Rauscher, M. , Ádám, A.L. , Wirtz, S. , Guggenheim, R. , Mendgen, K. & Deising, H.B. (1999) PR‐1 protein inhibits the differentiation of rust infection hyphae in leaves of acquired resistant broad bean. The Plant Journal, 19, 625–633. PubMed

Rivière, M.P. , Marais, A. , Ponchet, M. , Willats, W. & Galiana, E. (2008) Silencing of acidic pathogenesis‐related PR‐1 genes increases extracellular β‐(1→3)‐glucanase activity at the onset of tobacco defence reactions. Journal of Experimental Botany, 59, 1225–1239. PubMed

Saberianfar, R. & Menassa, R. (2017) Protein bodies: how the ER deals with high accumulation of recombinant proteins. Plant Biotechnology Journal, 15, 671–673. PubMed PMC

Santamaria, M. , Thomson, C.J. , Read, N.D. & Loake, G.J. (2001) The promoter of a basic PR1‐like gene, PubMed

Sarowar, S. , Young, J.K. , Eui, N.K. , Ki, D.K. , Byung, K.H. , Islam, R. et al. (2005) Overexpression of a pepper basic pathogenesis‐related protein 1 gene in tobacco plants enhances resistance to heavy metal and pathogen stresses. Plant Cell Reports, 24, 216–224. PubMed

Schindelin, J. , Arganda‐Carreras, I. , Frise, E. , Kaynig, V. , Longair, M. , Pietzsch, T. et al. (2012) Fiji: an open‐source platform for biological‐image analysis. Nature Methods, 9, 676–682. PubMed PMC

Sheng, J. , Olrichs, N.K. , Geerts, W.J. , Kaloyanova, D.V. & Helms, J.B. (2019) Metal ions and redox balance regulate distinct amyloid‐like aggregation pathways of GAPR‐1. Scientific Reports, 9, 15048. PubMed PMC

Sheng, J. , Olrichs, N.K. , Geerts, W.J. , Li, X. , Rehman, A.U. , Gadella, B.M. et al. (2019) Zinc binding regulates amyloid‐like aggregation of GAPR‐1. Bioscience Reports, 39, BSR20182345. PubMed PMC

Somssich, I.E. , Schmelzer, E. , Bollmann, J. & Hahlbrock, K. (1986) Rapid activation by fungal elicitor of genes encoding “pathogenesis‐related” proteins in cultured parsley cells. Proceedings of the National Academy of Sciences of the United States of America, 83, 2427–2430. PubMed PMC

Stintzi, A. , Heitz, T. , Prasad, V. , Wiedemann‐Merdinoglu, S. , Kauffmann, S. , Geoffroy, P. et al. (1993) Plant “pathogenesis‐related” proteins and their role in defense against pathogens. Biochimie, 75, 687–706. PubMed

Sung, Y.C. , Outram, M.A. , Breen, S. , Wang, C. , Dagvadorj, B. , Winterberg, B. et al. (2021) PR1‐mediated defence via C‐terminal peptide release is targeted by a fungal pathogen effector. New Phytologist, 229, 3467–3480. PubMed

Tao, K. , Waletich, J.R. , Wise, H. , Arredondo, F. & Tyler, B.M. (2019) Tethering of multi‐vesicular bodies and the tonoplast to the plasma membrane in plants. Frontiers in Plant Science, 10, 636. PubMed PMC

Tomczynska, I. , Stumpe, M. & Mauch, F. (2018) A conserved RxLR effector interacts with host RABA‐type GTPases to inhibit vesicle‐mediated secretion of antimicrobial proteins. The Plant Journal, 95, 187–203. PubMed

Vancová, M. , Bílý, T. , Nebesářová, J. , Grubhoffer, L. , Bonnet, S. , Park, Y. et al. (2019) Ultrastructural mapping of salivary gland innervation in the tick PubMed PMC

Watanabe, S. , Shimada, T.L. , Hiruma, K. & Takano, Y. (2013) Pathogen infection trial increases the secretion of proteins localized in the endoplasmic reticulum body of PubMed PMC

Webb, B. & Sali, A. (2016). Comparative protein structure modeling using MODELLER. Current Protocols in Protein Science, 2016, 5.6.1–5.6.37. PubMed

Woloshuk, C.P. , Meulenhoff, J.S. , Sela‐Buurlage, M. , van den Elzen, P.J.M. & Cornelissen, B.J.C. (1991) Pathogen‐induced proteins with inhibitory activity toward PubMed PMC

Wu, L. , Wu, H. , Chen, L. , Zhang, H. & Gao, X. (2017) Induction of systemic disease resistance in PubMed PMC

Yuan, J. & He, S.Y. (1996) The PubMed PMC

Zhang, W.J. , Pedersen, C. , Kwaaitaal, M. , Gregersen, P.L. , Mørch, S.M. , Hanisch, S. et al. (2012) Interaction of barley powdery mildew effector candidate CSEP0055 with the defence protein PR17c. Molecular Plant Pathology, 13, 1110–1119. PubMed PMC

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