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Actin from the apicomplexan Neospora caninum (NcACT) has different isoforms in 2D electrophoresis

Published online by Cambridge University Press:  06 June 2018

Luciana Baroni ,
Letícia Pollo-Oliveira ,
Albert JR Heck ,
AF Maarten Altelaar  and
Ana Patrícia Yatsuda Open the ORCID record for Ana Patrícia Yatsuda [Opens in a new window]
Luciana Baroni
Affiliation:
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. do Café, s/n, 14040-930, Ribeirão Preto, SP, Brazil
Letícia Pollo-Oliveira
Affiliation:
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. do Café, s/n, 14040-930, Ribeirão Preto, SP, Brazil
Albert JR Heck
Affiliation:
Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Centre for Biomolecular Research, Utrecht, University, Padualaan 8, Utrecht 3884 CH, The Netherlands
AF Maarten Altelaar
Affiliation:
Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Centre for Biomolecular Research, Utrecht, University, Padualaan 8, Utrecht 3884 CH, The Netherlands
Ana Patrícia Yatsuda
Affiliation:
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. do Café, s/n, 14040-930, Ribeirão Preto, SP, Brazil Departamento de Análises Clínicas, Bromatológicas e Toxicológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto 14040-930, Brazil
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Abstract

Apicomplexan parasites have unconventional actins that play a central role in important cellular processes such as apicoplast replication, motility of dense granules, endocytic trafficking and force generation for motility and host cell invasion. In this study, we investigated the actin of the apicomplexan Neospora caninum – a parasite associated with infectious abortion and neonatal mortality in livestock. Neospora caninum actin was detected and identified in two bands by one-dimensional (1D) western blot and in nine spots by the 2D technique. The mass spectrometry data indicated that N. caninum has at least nine different actin isoforms, possibly caused by post-translational modifications. In addition, the C4 pan-actin antibody detected specifically actin in N. caninum cellular extract. Extracellular N. caninum tachyzoites were treated with toxins that act on actin, jasplakinolide and cytochalasin D. Both substances altered the peripheric cytoplasmic localization of actin on tachyzoites. Our findings add complexity to the study of the apicomplexan actin in cellular processes, since the multiple functions of this important protein might be regulated by mechanisms involving post-translational modifications.

Keywords

Actinactin isoformsApicomplexacytochalasin D (CYTD)jasplakinolide (JAS)mass spectrometryNeospora caninum
Type
Research Article
Information
Parasitology , Volume 146 , Issue 1 , January 2019 , pp. 33 - 41
Copyright
Copyright © Cambridge University Press 2018 

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References

Alonso, A, Greenlee, M, Matts, J, Kline, J, Davis, KJ and Miller, RK (2015) Emerging roles of sumoylation in the regulation of actin, microtubules, intermediate filaments, and septins. Cytoskeleton (Hoboken) 72, 305–339.Google Scholar
Andreadaki, M, Morgan, RN, Deligianni, E, Kooij, TW, Santos, JM, Spanos, L, Matuschewski, K, Louis, C, Mair, GR and Siden-Kiamos, I (2014) Genetic crosses and complementation reveal essential functions for the Plasmodium stage-specific actin2 in sporogonic development. Cell Microbiology 16, 751–767.Google Scholar
Angrisano, F, Delves, MJ, Sturm, A, Mollard, V, McFadden, GI, Sinden, RE and Baum, J (2012 a) A GFP-actin reporter line to explore microfilament dynamics across the malaria parasite lifecycle. Molecular and Biochemical Parasitology 182, 93–96.Google Scholar
Angrisano, F, Riglar, DT, Sturm, A, Volz, JC, Delves, MJ, Zuccala, ES, Turnbull, L, Dekiwadia, C, Olshina, MA, Marapana, DS, Wong, W, Mollard, V, Bradin, CH, Tonkin, CJ, Gunning, PW, Ralph, SA, Whitchurch, CB, Sinden, RE, Cowman, AF, McFadden, GI and Baum, J (2012 b) Spatial localisation of actin filaments across developmental stages of the malaria parasite. PLoS ONE 7, e32188.Google Scholar
Baum, J, Papenfuss, AT, Baum, B, Speed, TP and Cowman, AF (2006) Regulation of apicomplexan actin-based motility. Nature Reviews Microbiology 4, 621–628.Google Scholar
Blanchoin, L, Boujemaa-Paterski, R, Sykes, C and Plastino, J (2014) Actin dynamics, architecture, and mechanics in cell motility. Physiological Reviews 94, 235–263.Google Scholar
Blom, N, Gammeltoft, S and Brunak, S (1999) Sequence and structure-based prediction of eukaryotic protein phosphorylation sites. Journal of Molecular Biology 294, 1351–1362.Google Scholar
Bradford, MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.Google Scholar
Bubb, M, Spector, I, Beyer, B and Fosen, K (2000) Effects of jasplakinolide on the kinetics of actin polymerization – an explanation for certain in vivo observations. Journal of Biological Chemistry 275, 5163–5170.Google Scholar
Bullard, B, Bell, J, Craig, R and Leonard, K (1985) Arthrin: a new actin-like protein in insect flight muscle. Journal of Molecular Biology 182, 443–454.Google Scholar
Burgess, S, Walker, M, Knight, P, Sparrow, J, Schmitz, S, Offer, G, Bullard, B, Leonard, K, Holt, J and Trinick, J (2004) Structural studies of arthrin: monoubiquitinated actin. Journal of Molecular Biology 341, 1161–1173.Google Scholar
Castro, J, Ott, C, Jung, T, Grune, T and Almeida, H (2012) Carbonylation of the cytoskeletal protein actin leads to aggregate formation. Free Radical Biology and Medicine 53, 916–925.Google Scholar
Cevallos, AM, Segura-Kato, YX, Merchant-Larios, H, Manning-Cela, R, Alberto Hernández-Osorio, L, Márquez-Dueñas, C, Ambrosio, JR, Reynoso-Ducoing, O and Hernández, R (2011) Trypanosoma cruzi: multiple actin isovariants are observed along different developmental stages. Experimental Parasitology 127, 249–259.Google Scholar
Cooper, JA (1987) Effects of cytochalasin and phalloidin on actin. Journal of Cell Biology 105, 1473–1478.Google Scholar
Deligianni, E, Morgan, RN, Bertuccini, L, Kooij, TW, Laforge, A, Nahar, C, Poulakakis, N, Schüler, H, Louis, C, Matuschewski, K and Siden-Kiamos, I (2011) Critical role for a stage-specific actin in male exflagellation of the malaria parasite. Cell Microbiology 13, 1714–1730.Google Scholar
Deng, W, Wang, C, Zhang, Y, Xu, Y, Zhang, S, Liu, Z and Xue, Y (2016) GPS-PAIL: prediction of lysine acetyltransferase-specific modification sites from protein sequences. Scientific Reports 6, 39787.Google Scholar
Dobrowolski, JM and Sibley, LD (1996) Toxoplasma invasion of mammalian cells is powered by the actin cytoskeleton of the parasite. Cell 84, 933–939.Google Scholar
Dobrowolski, J, Niesman, I and Sibley, L (1997) Actin in the parasite Toxoplasma gondii is encoded by a single copy gene, ACT1 and exists primarily in a globular form. Cell Motility and the Cytoskeleton 37, 253–262.Google Scholar
Drewry, LL and Sibley, LD (2015) Toxoplasma actin is required for efficient host cell invasion. MBio 6, e00557.Google Scholar
Dubey, J and Schares, G (2011) Neosporosis in animals – the last five years. Veterinary Parasitology 180, 90–108.Google Scholar
Dubey, J, Hattel, A, Lindsay, D and Topper, M (1988) Neonatal Neospora caninum infection in dogs – isolation of the causative agent and experimental transmission. Journal of the American Veterinary Medical Association 193, 1259–1263.Google Scholar
Field, SJ, Pinder, JC, Clough, B, Dluzewski, AR, Wilson, RJ and Gratzer, WB (1993) Actin in the merozoite of the malaria parasite, Plasmodium falciparum. Cell Motility Cytoskeleton 25, 43–48.Google Scholar
Frölich, S and Wallach, M (2015) F-actin distribution and function during sexual development in Eimeria maxima. Parasitology 142, 855–864.Google Scholar
Gajria, B, Bahl, A, Brestelli, J, Dommer, J, Fischer, S, Gao, X, Heiges, M, Iodice, J, Kissinger, JC, Mackey, AJ, Pinney, DF, Roos, DS, Stoeckert, CJ, Wang, H and Brunk, BP (2008) ToxoDB: an integrated Toxoplasma gondii database resource. Nucleic Acids Research 36, D553–D556.Google Scholar
González-López, L, Carballar-Lejarazú, R, Arrevillaga Boni, G, Cortés-Martínez, L, Cázares-Raga, FE, Trujillo-Ocampo, A, Rodríguez, MH, James, AA and Hernández-Hernández, FC (2017) Lys48 ubiquitination during the intraerythrocytic cycle of the rodent malaria parasite, Plasmodium chabaudi. PLoS ONE 12, e0176533.Google Scholar
Gordon, JL and Sibley, LD (2005) Comparative genome analysis reveals a conserved family of actin-like proteins in apicomplexan parasites. BMC Genomics 6, 179.Google Scholar
Gordon, JL, Buguliskis, JS, Buske, PJ and Sibley, LD (2010) Actin-like protein 1 (ALP1) is a component of dynamic, high molecular weight complexes in Toxoplasma gondii. Cytoskeleton (Hoboken) 67, 23–31.Google Scholar
Gu, L, Zhang, H, Chen, Q and Chen, J (2003) Calyculin A-induced actin phosphorylation and depolymerization in renal epithelial cells. Cell Motility and the Cytoskeleton 54, 286–295.Google Scholar
Gupta, CM, Thiyagarajan, S and Sahasrabuddhe, AA (2015) Unconventional actins and actin-binding proteins in human protozoan parasites. International Journal of Parasitology 45, 435–447.Google Scholar
Haase, S, Zimmermann, D, Olshina, MA, Wilkinson, M, Fisher, F, Tan, YH, Stewart, RJ, Tonkin, CJ, Wong, W, Kovar, DR and Baum, J (2015) Disassembly activity of actin-depolymerizing factor (ADF) is associated with distinct cellular processes in apicomplexan parasites. Molecular Biology of the Cell 26, 3001–3012.Google Scholar
Hasan, MA, Li, J, Ahmad, S and Molla, MK (2017) PredCar-site: carbonylation sites prediction in proteins using support vector machine with resolving data imbalanced issue. Analytical Biochemistry 525, 107–113.Google Scholar
Heaslip, AT, Nelson, SR and Warshaw, DM (2016) Dense granule trafficking in Toxoplasma gondii requires a unique class 27 myosin and actin filaments. Molecular Biology of the Cell 27, 2080–2089.Google Scholar
Heintzelman, MB (2015) Gliding motility in apicomplexan parasites. Seminars in Cell & Developmental Biology 46, 135–142.Google Scholar
Kang, H, Bradley, MJ, Elam, WA and De La Cruz, EM (2013) Regulation of actin by ion-linked equilibria. Biophysical Journal 105, 2621–2628.Google Scholar
Karakozova, M, Kozak, M, Wong, C, Bailey, A, Yates, J, Mogilner, A, Zebroski, H and Kashina, A (2006) Arginylation of beta-actin regulates actin cytoskeleton and cell motility. Science 313, 192–196.Google Scholar
Kishi, Y, Clements, C, Mahadeo, D, Cotter, D and Sameshima, M (1998) High levels of actin tyrosine phosphorylation: correlation with the dormant state of dictyostelium spores. Journal of Cell Science 111, 2923–2932.Google Scholar
Kübler, E and Riezman, H (1993) Actin and fimbrin are required for the internalization step of endocytosis in yeast. EMBO Journal 12, 2855–2862.Google Scholar
Kudryashov, DS and Reisler, E (2013) ATP and ADP actin states. Biopolymers 99, 245–256.Google Scholar
Kumpula, EP, Pires, I, Lasiwa, D, Piirainen, H, Bergmann, U, Vahokoski, J and Kursula, I (2017) Apicomplexan actin polymerization depends on nucleation. Scientific Reports 7, 12137.Google Scholar
Lappalainen, P (2016) Actin-binding proteins: the long road to understanding the dynamic landscape of cellular actin networks. Molecular Biology of the Cell 27, 2519–2522.Google Scholar
Lee, E, Kim, J, Shin, Y, Shin, G, Suh, M, Kim, D, Kim, Y, Kim, G and Jung, T (2003) Establishment of a two-dimensional electrophoresis map for Neospora caninum tachyzoites by proteomics. Proteomics 3, 2339–2350.Google Scholar
Lessard, JL (1988) Two monoclonal antibodies to actin: one muscle selective and one generally reactive. Cell Motility and the Cytoskeleton 10, 349–362.Google Scholar
Lu, J, Katano, T, Okuda-Ashitaka, E, Oishi, Y, Urade, Y and Ito, S (2009) Involvement of S-nitrosylation of actin in inhibition of neurotransmitter release by nitric oxide. Molecular Pain 5, 58.Google Scholar
Matsudaira, P (1991) Modular organization of actin crosslinking proteins. Trends in Biochemical Sciences 16, 87–92.Google Scholar
Mizuno, Y, Makioka, A, Kawazu, S, Kano, S, Kawai, S, Akaki, M, Aikawa, M and Ohtomo, H (2002) Effect of jasplakinolide on the growth, invasion, and actin cytoskeleton of Plasmodium falciparum. Parasitology Research 88, 844–848.Google Scholar
Nyman, T, Schuler, H, Korenbaum, E, Schutt, C, Karlsson, R and Lindberg, U (2002) The role of MeH73 in actin polymerization and ATP hydrolysis. Journal of Molecular Biology 317, 577–589.Google Scholar
Opitz, C and Soldati, D (2002) ‘The glideosome’: a dynamic complex powering gliding motion and host cell invasion by Toxoplasma gondii. Molecular Microbiology 45, 597–604.Google Scholar
Pereira, LM, Candido-Silva, JA, De Vries, E and Yatsuda, AP (2011) A new thrombospondin-related anonymous protein homologue in Neospora caninum (NcMIC2-like1). Parasitology 138, 287–297.Google Scholar
Periz, J, Whitelaw, J, Harding, C, Gras, S, Del Rosario Minina, MI, Latorre-Barragan, F, Lemgruber, L, Reimer, MA, Insall, R, Heaslip, A and Meissner, M (2017) Toxoplasma gondii F-actin forms an extensive filamentous network required for material exchange and parasite maturation. Elife 6, e24119.Google Scholar
Perrin, BJ and Ervasti, JM (2010) The actin gene family: function follows isoform. Cytoskeleton (Hoboken) 67, 630–634.Google Scholar
Pollard, T and Borisy, G (2003) Cellular motility driven by assembly and disassembly of actin filaments. Cell 112, 453–465.Google Scholar
Pollo-Oliveira, L, Post, H, Acencio, ML, Lemke, N, van den Toorn, H, Tragante, V, Heck, AJ, Altelaar, AF and Yatsuda, AP (2013) Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics. Parasites & Vectors 6, 335.Google Scholar
Reichel, M, Ayanegui-Alcerreca, M, Gondim, L and Ellis, J (2013) What is the global economic impact of Neospora caninum in cattle – The billion dollar question. International Journal for Parasitology 43, 133–142.Google Scholar
Rubenstein, PA and Martin, DJ (1983) NH2-terminal processing of Drosophila melanogaster actin. Sequential removal of two amino acids. Journal of Biological Chemistry 258, 11354–11360.Google Scholar
Sahoo, N, Beatty, W, Heuser, J, Sept, D and Sibley, L (2006) Unusual kinetic and structural properties control rapid assembly and turnover of actin in the parasite Toxoplasma gondii. Molecular Biology of the Cell 17, 895–906.Google Scholar
Sakai, J, Li, J, Subramanian, KK, Mondal, S, Bajrami, B, Hattori, H, Jia, Y, Dickinson, BC, Zhong, J, Ye, K, Chang, CJ, Ho, YS, Zhou, J and Luo, HR (2012) Reactive oxygen species-induced actin glutathionylation controls actin dynamics in neutrophils. Immunity 37, 1037–1049.Google Scholar
Schafer, DA and Schroer, TA (1999) Actin-related proteins. Annual Review of Cell Developmental Biology 15, 341–363.Google Scholar
Shaw, M and Tilney, L (1999) Induction of an acrosomal process in Toxoplasma gondii: visualization of actin filaments in a protozoan parasite. Proceedings of the National Academy of Sciences of the USA 96, 9095–9099.Google Scholar
Siden-Kiamos, I, Pinder, JC and Louis, C (2006) Involvement of actin and myosins in Plasmodium berghei ookinete motility. Molecular and Biochemical Parasitology 150, 308–317.Google Scholar
Skillman, KM, Ma, CI, Fremont, DH, Diraviyam, K, Cooper, JA, Sept, D and Sibley, LD (2013) The unusual dynamics of parasite actin result from isodesmic polymerization. Nature Communications 4, 2285.Google Scholar
Slajcherová, K, Fišerová, J, Fischer, L and Schwarzerová, K (2012) Multiple actin isotypes in plants: diverse genes for diverse roles? Frontiers in Plant Science 3, 226.Google Scholar
Smythe, WA, Joiner, KA and Hoppe, HC (2008) Actin is required for endocytic trafficking in the malaria parasite Plasmodium falciparum. Cellular Microbiology 10, 452–464.Google Scholar
Soldati, D and Meissner, M (2004) Toxoplasma as a novel system for motility. Current Opinion in Cell Biology 16, 32–40.Google Scholar
Tardieux, I and Baum, J (2016) Reassessing the mechanics of parasite motility and host-cell invasion. The Journal of Cell Biology 214, 507–515.Google Scholar
Terman, JR and Kashina, A (2013) Post-translational modification and regulation of actin. Current Opinion in Cell Biology 25, 30–38.Google Scholar
The UniProt Consortium (2017) Uniprot: the universal protein knowledgebase. Nucleic Acids Research 45, D158–D169.Google Scholar
Vahokoski, J, Bhargav, SP, Desfosses, A, Andreadaki, M, Kumpula, EP, Martinez, SM, Ignatev, A, Lepper, S, Frischknecht, F, Sidén-Kiamos, I, Sachse, C and Kursula, I (2014) Structural differences explain diverse functions of Plasmodium actins. PLoS Pathogens 10, e1004091.Google Scholar
Wesseling, JG, Snijders, PJ, van Someren, P, Jansen, J, Smits, MA and Schoenmakers, JG (1989) Stage-specific expression and genomic organization of the actin genes of the malaria parasite Plasmodium falciparum. Molecular and Biochemical Parasitology 35, 167–176.Google Scholar
Wetzel, DM, Håkansson, S, Hu, K, Roos, D and Sibley, LD (2003) Actin filament polymerization regulates gliding motility by apicomplexan parasites. Molecular Biology of the Cell 14, 396–406.Google Scholar
Whitelaw, JA, Latorre-Barragan, F, Gras, S, Pall, GS, Leung, JM, Heaslip, A, Egarter, S, Andenmatten, N, Nelson, SR, Warshaw, DM, Ward, GE and Meissner, M (2017) Surface attachment, promoted by the actomyosin system of Toxoplasma gondii is important for efficient gliding motility and invasion. BMC Biology 15, 1.Google Scholar
Xia, D, Sanderson, SJ, Jones, AR, Prieto, JH, Yates, JR, Bromley, E, Tomley, FM, Lal, K, Sinden, RE, Brunk, BP, Roos, DS and Wastling, JM (2008) The proteome of Toxoplasma gondii: integration with the genome provides novel insights into gene expression and annotation. Genome Biology 9, R116.Google Scholar
Xue, Y, Liu, Z, Gao, X, Jin, C, Wen, L, Yao, X and Ren, J (2010) GPS-SNO: computational prediction of protein S-nitrosylation sites with a modified GPS algorithm. PLoS ONE 5, e11290.Google Scholar
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