Hostname: page-component-cd9895bd7-dk4vv Total loading time: 0 Render date: 2024-12-26T13:00:55.077Z Has data issue: false hasContentIssue false
  • English
  • Français

Neospora caninum cytoplasmic dynein LC8 light chain 2 (NcDYNLL2) is differentially produced by pathogenically distinct isolates and regulates the host immune response

Published online by Cambridge University Press:  18 December 2018

Lili Cao ,
Raymond Fetterer ,
Guanggang Qu ,
Xichen Zhang  and
Wenbin Tuo Open the ORCID record for Wenbin Tuo [Opens in a new window]
Lili Cao
Affiliation:
Jilin Academy of Animal Science and Veterinary Medicine, and College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China Animal Parasitic Diseases Laboratory, Agricultural Research Service, USDA, Beltsville, MD 20705, USA
Raymond Fetterer
Affiliation:
Animal Parasitic Diseases Laboratory, Agricultural Research Service, USDA, Beltsville, MD 20705, USA
Guanggang Qu
Affiliation:
Animal Parasitic Diseases Laboratory, Agricultural Research Service, USDA, Beltsville, MD 20705, USA
Xichen Zhang
Affiliation:
Jilin Academy of Animal Science and Veterinary Medicine, and College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Wenbin Tuo*
Affiliation:
Animal Parasitic Diseases Laboratory, Agricultural Research Service, USDA, Beltsville, MD 20705, USA
*
Author for correspondence: Wenbin Tuo, E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Neospora caninum is the causative agent of bovine neosporosis. A N. caninum cytoplasmic dynein LC8 light chain (NcDYNLL) protein was characterized in this study. Cytoplasmic dyneins, including DYNLLs, belong to the microtubule minus-end-directed motor proteins and are involved in many cellular processes. Previous microarray studies revealed that NcDYNLL was downregulated in the non-pathogenic clone, Ncts-8, when compared with the wild-type NC1 isolate. The present study showed that DYNLLs from different species are highly conserved (>85% identity), and the NcDYNLL belongs to the DYNLL2 family. NcDYNLL2 and Toxoplasma gondii DYNLL2 have identical amino acid sequences, although they are slightly divergent at the genetic level (89% identity). NcDYNLL2 was cloned and expressed in Escherichia coli and purified. NcDYNLL2 was identified in soluble and insoluble fractions of tachyzoite lysate. As expected, soluble NcDYNLL2 was lower in the Ncts-8 lysate when compared with that of NC1 isolate. NcDYNLL2 release by the tachyzoites was low; however, it was increased when tachyzoites were treated with either calcium ionophore or ethanol. The data indicate that NcDYNLL2 may be actively secreted at low levels, but the secretion was upregulated by agents that also augment microneme protein secretions. Immunostaining of NcDYNLL2 in isolated and intracellular Neospora tachyzoites showed a diffuse distribution pattern. Furthermore, rNcDYNLL2 was internalized by the host immune cells and stimulated tumour necrosis factor-α) and interleukin-12 (IL-12) production by murine dendritic cells. Taken together, these results suggest that NcDYNLL2 is a secretory protein that cross-regulates host immunity.

Keywords

Dynein LC8 light chain 2DYNLL2Neospora caninumToxoplasma gondii
Type
Research Article
Information
Parasitology , Volume 146 , Issue 5 , April 2019 , pp. 588 - 595
Check for updates
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. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aida, Y and Pabst, MJ (1990) Removal of endotoxin from protein solutions by phase separation using Triton X-114. Journal of Immunological Methods 132, 191–195.Google Scholar
Barbar, E (2008) Dynein light chain LC8 is a dimerization hub essential in diverse protein networks. Biochemistry 47, 503–508.Google Scholar
Carruthers, VB and Tomley, FM (2008) Microneme proteins in apicomplexans. Sub-Cellular Biochemistry 47, 33–45.Google Scholar
Day, CL, Puthalakath, H, Skea, G, Strasser, A, Barsukov, I, Lian, LY, Huang, DC and Hinds, MG (2004) Localization of dynein light chains 1 and 2 and their pro-apoptotic ligands. Biochemical Journal 377, 597–605.Google Scholar
Dreier, KJ, Stewarter, LW, Kerlin, RL, Ritter, DM and Brake, DA (1999) Phenotypic characterisation of a Neospora caninum temperature-sensitive strain in normal and immunodeficient mice. International Journal for Parasitology 29, 1627–1634.Google Scholar
Dubey, JP, Buxton, D and Wouda, W (2006) Pathogenesis of bovine neosporosis. Journal of Comparative Pathology 134, 267–289.Google Scholar
Feng, X, Zhang, N and Tuo, W (2010) Neospora caninum tachyzoite- and antigen-stimulated cytokine production by bone marrow-derived dendritic cells and spleen cells of naive BALB/c mice. Journal of Parasitology 96, 717–723.Google Scholar
Githui, EK, De Villiers, EP and McArthur, AG (2009) Plasmodium possesses dynein light chain classes that are unique and conserved across species. Infection Genetics and Evolution 9, 337–343.Google Scholar
Jaffrey, SR and Snyder, SH (1996) PIN: an associated protein inhibitor of neuronal nitric oxide synthase. Science 274, 774–777.Google Scholar
Jeong, W, Chang, TS, Boja, ES, Fales, HM and Rhee, SG (2004) Roles of TRP14, a thioredoxin-related protein in tumor necrosis factor-alpha signaling pathways. Journal of Biological Chemistry 279, 3151–3159.Google Scholar
Jeong, W, Jung, Y, Kim, H, Park, SJ and Rhee, SG (2009) Thioredoxin-related protein 14, a new member of the thioredoxin family with disulfide reductase activity: implication in the redox regulation of TNF-alpha signaling. Free Radical Biology & Medicine 47, 1294–1303.Google Scholar
Jung, Y, Kim, H, Min, SH, Rhee, SG and Jeong, W (2008) Dynein light chain LC8 negatively regulates NF-kappaB through the redox-dependent interaction with IkappaBalpha. Journal of Biological Chemistry 283, 23863–23871.Google Scholar
Jurado, S, Gleeson, K, O'Donnell, K, Izon, DJ, Walkley, CR, Strasser, A, Tarlinton, DM and Heierhorst, J (2012) The zinc-finger protein ASCIZ regulates B cell development via DYNLL1 and Bim. Journal of Experimental Medicine 209, 1629–1639.Google Scholar
Li, RW and Tuo, W (2011) Neospora caninum: comparative gene expression profiling of Neospora caninum wild type and a temperature sensitive clone. Experimental Parasitology 129, 346–354.Google Scholar
Lindsay, DS, Lenz, SD, Blagburn, BL and Brake, DA (1999) Characterization of temperature-sensitive strains of Neospora caninum in mice. Journal of Parasitology 85, 64–67.Google Scholar
Naisbitt, S, Valtschanoff, J, Allison, DW, Sala, C, Kim, E, Craig, AM, Weinberg, RJ and Sheng, M (2000) Interaction of the postsynaptic density-95/guanylate kinase domain-associated protein complex with a light chain of myosin-V and dynein. Journal of Neuroscience 20, 4524–4534.Google Scholar
Pfister, KK, Fisher, EM, Gibbons, IR, Hays, TS, Holzbaur, EL, McIntosh, JR, Porter, ME, Schroer, TA, Vaughan, KT, Witman, GB, King, SM and Vallee, RB (2005) Cytoplasmic dynein nomenclature. Journal of Cell Biology 171, 411–413.Google Scholar
Pfister, KK, Shah, PR, Hummerich, H, Russ, A, Cotton, J, Annuar, AA, King, SM and Fisher, EM (2006) Genetic analysis of the cytoplasmic dynein subunit families. PLoS Genetics 2, e1.Google Scholar
Puthalakath, H, Huang, DC, O'Reilly, LA, King, SM and Strasser, A (1999) The proapoptotic activity of the Bcl-2 family member Bim is regulated by interaction with the dynein motor complex. Molecular Cell 3, 287–296.Google Scholar
Puthalakath, H, Villunger, A, O'Reilly, LA, Beaumont, JG, Coultas, L, Cheney, RE, Huang, DC and Strasser, A (2001) Bmf: a proapoptotic BH3-only protein regulated by interaction with the myosin V actin motor complex, activated by anoikis. Science 293, 1829–1832.Google Scholar
Qu, G, Fetterer, R, Jenkins, M, Leng, L, Shen, Z, Murphy, C, Han, W, Bucala, R and Tuo, W (2013) Characterization of Neospora caninum macrophage migration inhibitory factor. Experimental Parasitology 135, 246–256.Google Scholar
Qu, G, Fetterer, R, Leng, L, Du, X, Zarlenga, D, Shen, Z, Han, W, Bucala, R and Tuo, W (2014) Ostertagia ostertagi macrophage migration inhibitory factor is present in all developmental stages and may cross-regulate host functions through interaction with the host receptor. International Journal for Parasitology 44, 355–367.Google Scholar
Qureshi, BM, Hofmann, NE, Arroyo-Olarte, RD, Nickl, B, Hoehne, W, Jungblut, PR, Lucius, R, Scheerer, P and Gupta, N (2013) Dynein light chain 8a of Toxoplasma gondii, a unique conoid-localized beta-strand-swapped homodimer, is required for an efficient parasite growth. FASEB Journal 27, 1034–1047.Google Scholar
Raux, H, Flamand, A and Blondel, D (2000) Interaction of the rabies virus P protein with the LC8 dynein light chain. Journal of Virology 74, 10212–10216.Google Scholar
Rayala, SK, den, HP, Balasenthil, S, Yang, Z, Broaddus, RR and Kumar, R (2005) Functional regulation of oestrogen receptor pathway by the dynein light chain 1. EMBO Reports 6, 538–544.Google Scholar
Ritter, DM, Kerlin, R, Sibert, G and Brake, D (2002) Immune factors influencing the course of infection with Neospora caninum in the murine host. Journal of Parasitology 88, 271–280.Google Scholar
Schnorrer, F, Bohmann, K and Nusslein-Volhard, C (2000) The molecular motor dynein is involved in targeting swallow and bicoid RNA to the anterior pole of Drosophila oocytes. Nature Cell Biology 2, 185–190.Google Scholar
Stober, CB, Lange, UG, Roberts, MT, Gilmartin, B, Francis, R, Almeida, R, Peacock, CS, McCann, S and Blackwell, JM (2006) From genome to vaccines for leishmaniasis: screening 100 novel vaccine candidates against murine Leishmania major infection. Vaccine 24, 2602–2616.Google Scholar
Tuo, W, Fetterer, R, Jenkins, M and Dubey, JP (2005) Identification and characterization of Neospora caninum cyclophilin that elicits gamma interferon production. Infection and Immunity 73, 5093–5100.Google Scholar
Tuo, W, Zhao, Y, Zhu, D and Jenkins, MC (2011) Immunization of female BALB/c mice with Neospora cyclophilin and/or NcSRS2 elicits specific antibody response and prevents against challenge infection by Neospora caninum. Vaccine 29, 2392–2399.Google Scholar
Vadlamudi, RK, Bagheri-Yarmand, R, Yang, Z, Balasenthil, S, Nguyen, D, Sahin, AA, den, HP and Kumar, R (2004) Dynein light chain 1, a p21-activated kinase 1-interacting substrate, promotes cancerous phenotypes. Cancer Cell 5, 575–585.Google Scholar
Wilkes, DE, Rajagopalan, V, Chan, CW, Kniazeva, E, Wiedeman, AE and Asai, DJ (2007) Dynein light chain family in Tetrahymena thermophila. Cell Motility and the Cytoskeleton 64, 82–96.Google Scholar
Wilson, MJ, Salata, MW, Susalka, SJ and Pfister, KK (2001) Light chains of mammalian cytoplasmic dynein: identification and characterization of a family of LC8 light chains. Cell Motility and the Cytoskeleton 49, 229–240.Google Scholar
Yang, P, Diener, DR, Rosenbaum, JL and Sale, WS (2001) Localization of calmodulin and dynein light chain LC8 in flagellar radial spokes. Journal of Cell Biology 153, 1315–1326.Google Scholar
Yin, J, Qu, G, Cao, L, Li, Q, Fetterer, R, Feng, X, Liu, Q, Wang, G, Qi, D, Zhang, X, Miramontes, E, Jenkins, M, Zhang, N and Tuo, W (2012) Characterization of Neospora caninum microneme protein 10 (NcMIC10) and its potential use as a diagnostic marker for neosporosis. Veterinary Parasitology 187, 28–35.Google Scholar