Skip to main content Accessibility help
×
Home
Hostname: page-component-559fc8cf4f-lzpzj Total loading time: 0.415 Render date: 2021-03-04T10:52:59.074Z Has data issue: true Feature Flags: { "shouldUseShareProductTool": true, "shouldUseHypothesis": true, "isUnsiloEnabled": true, "metricsAbstractViews": false, "figures": false, "newCiteModal": false, "newCitedByModal": true }

Article contents

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

Published online by Cambridge University Press:  06 June 2018

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
Corresponding
E-mail address:

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.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below.

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, 305339.CrossRefGoogle ScholarPubMed
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, 751767.CrossRefGoogle ScholarPubMed
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, 9396.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
Baum, J, Papenfuss, AT, Baum, B, Speed, TP and Cowman, AF (2006) Regulation of apicomplexan actin-based motility. Nature Reviews Microbiology 4, 621628.CrossRefGoogle ScholarPubMed
Blanchoin, L, Boujemaa-Paterski, R, Sykes, C and Plastino, J (2014) Actin dynamics, architecture, and mechanics in cell motility. Physiological Reviews 94, 235263.CrossRefGoogle ScholarPubMed
Blom, N, Gammeltoft, S and Brunak, S (1999) Sequence and structure-based prediction of eukaryotic protein phosphorylation sites. Journal of Molecular Biology 294, 13511362.CrossRefGoogle ScholarPubMed
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, 248254.CrossRefGoogle ScholarPubMed
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, 51635170.CrossRefGoogle ScholarPubMed
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, 443454.CrossRefGoogle ScholarPubMed
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, 11611173.CrossRefGoogle ScholarPubMed
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, 916925.CrossRefGoogle ScholarPubMed
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, 249259.CrossRefGoogle ScholarPubMed
Cooper, JA (1987) Effects of cytochalasin and phalloidin on actin. Journal of Cell Biology 105, 14731478.CrossRefGoogle ScholarPubMed
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, 17141730.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
Dobrowolski, JM and Sibley, LD (1996) Toxoplasma invasion of mammalian cells is powered by the actin cytoskeleton of the parasite. Cell 84, 933939.CrossRefGoogle ScholarPubMed
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, 253262.3.0.CO;2-7>CrossRefGoogle Scholar
Drewry, LL and Sibley, LD (2015) Toxoplasma actin is required for efficient host cell invasion. MBio 6, e00557.CrossRefGoogle ScholarPubMed
Dubey, J and Schares, G (2011) Neosporosis in animals – the last five years. Veterinary Parasitology 180, 90108.CrossRefGoogle ScholarPubMed
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, 12591263.Google ScholarPubMed
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, 4348.CrossRefGoogle ScholarPubMed
Frölich, S and Wallach, M (2015) F-actin distribution and function during sexual development in Eimeria maxima. Parasitology 142, 855864.CrossRefGoogle ScholarPubMed
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, D553D556.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
Gordon, JL and Sibley, LD (2005) Comparative genome analysis reveals a conserved family of actin-like proteins in apicomplexan parasites. BMC Genomics 6, 179.CrossRefGoogle ScholarPubMed
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, 2331.Google ScholarPubMed
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, 286295.CrossRefGoogle ScholarPubMed
Gupta, CM, Thiyagarajan, S and Sahasrabuddhe, AA (2015) Unconventional actins and actin-binding proteins in human protozoan parasites. International Journal of Parasitology 45, 435447.CrossRefGoogle ScholarPubMed
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, 30013012.CrossRefGoogle ScholarPubMed
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, 107113.CrossRefGoogle ScholarPubMed
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, 20802089.CrossRefGoogle ScholarPubMed
Heintzelman, MB (2015) Gliding motility in apicomplexan parasites. Seminars in Cell & Developmental Biology 46, 135142.CrossRefGoogle ScholarPubMed
Kang, H, Bradley, MJ, Elam, WA and De La Cruz, EM (2013) Regulation of actin by ion-linked equilibria. Biophysical Journal 105, 26212628.CrossRefGoogle ScholarPubMed
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, 192196.CrossRefGoogle ScholarPubMed
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, 29232932.Google ScholarPubMed
Kübler, E and Riezman, H (1993) Actin and fimbrin are required for the internalization step of endocytosis in yeast. EMBO Journal 12, 28552862.CrossRefGoogle Scholar
Kudryashov, DS and Reisler, E (2013) ATP and ADP actin states. Biopolymers 99, 245256.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
Lappalainen, P (2016) Actin-binding proteins: the long road to understanding the dynamic landscape of cellular actin networks. Molecular Biology of the Cell 27, 25192522.CrossRefGoogle 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, 23392350.CrossRefGoogle ScholarPubMed
Lessard, JL (1988) Two monoclonal antibodies to actin: one muscle selective and one generally reactive. Cell Motility and the Cytoskeleton 10, 349362.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
Matsudaira, P (1991) Modular organization of actin crosslinking proteins. Trends in Biochemical Sciences 16, 8792.CrossRefGoogle ScholarPubMed
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, 844848.CrossRefGoogle ScholarPubMed
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, 577589.CrossRefGoogle ScholarPubMed
Opitz, C and Soldati, D (2002) ‘The glideosome’: a dynamic complex powering gliding motion and host cell invasion by Toxoplasma gondii. Molecular Microbiology 45, 597604.CrossRefGoogle ScholarPubMed
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, 287297.CrossRefGoogle 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.CrossRefGoogle ScholarPubMed
Perrin, BJ and Ervasti, JM (2010) The actin gene family: function follows isoform. Cytoskeleton (Hoboken) 67, 630634.CrossRefGoogle ScholarPubMed
Pollard, T and Borisy, G (2003) Cellular motility driven by assembly and disassembly of actin filaments. Cell 112, 453465.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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, 133142.CrossRefGoogle ScholarPubMed
Rubenstein, PA and Martin, DJ (1983) NH2-terminal processing of Drosophila melanogaster actin. Sequential removal of two amino acids. Journal of Biological Chemistry 258, 1135411360.Google ScholarPubMed
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, 895906.CrossRefGoogle ScholarPubMed
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, 10371049.CrossRefGoogle ScholarPubMed
Schafer, DA and Schroer, TA (1999) Actin-related proteins. Annual Review of Cell Developmental Biology 15, 341363.CrossRefGoogle ScholarPubMed
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, 90959099.CrossRefGoogle 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, 308317.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
Smythe, WA, Joiner, KA and Hoppe, HC (2008) Actin is required for endocytic trafficking in the malaria parasite Plasmodium falciparum. Cellular Microbiology 10, 452464.Google ScholarPubMed
Soldati, D and Meissner, M (2004) Toxoplasma as a novel system for motility. Current Opinion in Cell Biology 16, 3240.CrossRefGoogle ScholarPubMed
Tardieux, I and Baum, J (2016) Reassessing the mechanics of parasite motility and host-cell invasion. The Journal of Cell Biology 214, 507515.CrossRefGoogle ScholarPubMed
Terman, JR and Kashina, A (2013) Post-translational modification and regulation of actin. Current Opinion in Cell Biology 25, 3038.CrossRefGoogle ScholarPubMed
The UniProt Consortium (2017) Uniprot: the universal protein knowledgebase. Nucleic Acids Research 45, D158D169.CrossRefGoogle 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.CrossRefGoogle ScholarPubMed
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, 167176.CrossRefGoogle ScholarPubMed
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, 396406.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed

Baroni et al. supplementary material

Tables S5-S8

File 30 KB

Baroni et al. supplementary material

Table S1

File 20 KB

Baroni et al. supplementary material

Table S2

File 22 KB

Baroni et al. supplementary material

Table S3

File 35 KB

Baroni et al. supplementary material

Table S4

File 44 KB

Altmetric attention score

Full text views

Full text views reflects PDF downloads, PDFs sent to Google Drive, Dropbox and Kindle and HTML full text views.

Total number of HTML views: 34
Total number of PDF views: 164 *
View data table for this chart

* Views captured on Cambridge Core between 06th June 2018 - 4th March 2021. This data will be updated every 24 hours.

Send article to Kindle

To send this article to your Kindle, first ensure no-reply@cambridge.org is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about sending to your Kindle. Find out more about sending to your Kindle.

Note you can select to send to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be sent to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

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

Send article to Dropbox

To send this article to your Dropbox account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Dropbox.

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

Send article to Google Drive

To send this article to your Google Drive account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Google Drive.

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

Reply to: Submit a response


Your details


Conflicting interests

Do you have any conflicting interests? *