Expression and Identification of a Novel Spore Wall Protein in Microsporidian Nosema bombycis.
IFA
microsporidia
tandem repeats
Journal
The Journal of eukaryotic microbiology
ISSN: 1550-7408
Titre abrégé: J Eukaryot Microbiol
Pays: United States
ID NLM: 9306405
Informations de publication
Date de publication:
11 2020
11 2020
Historique:
received:
10
12
2019
revised:
07
07
2020
accepted:
14
07
2020
pubmed:
24
7
2020
medline:
22
6
2021
entrez:
24
7
2020
Statut:
ppublish
Résumé
Microsporidia are a group of obligate intracellular parasites causing significant disease in human beings and economically important animals. Though a few spore wall proteins (SWPs) have now been identified in these intriguing species, the information on SWPs remains too little to elucidate the spore wall formation mechanisms of microsporidia. It has been well described that numerous proteins with tandem repeats tend to be localized on the cell wall of fungi and parasites. Previously, by scanning the proteins with tandem repeats in microsporidian Nosema bombycis, we obtained 83 candidate SWPs based on whether those proteins possess a signal peptide and/or transmembrane domain. Here, we further characterized a candidate protein (EOB13250) with three tandem repeats in the N-terminal region and a transmembrane domain in C-terminus of N. bombycis. Sequence analysis showed that the tandem repeat domain of EOB13250 was species-specific for this parasite. RT-PCR indicated that the expression of the gene encoding this protein started on the fourth day postinfection. After cloned and expressed in Escherichia coli, a polyclone antibody against the recombinant EOB13250 protein was prepared. Western blotting demonstrated this protein exist in N. bombycis. Immunofluorescence analysis (IFA) and immunoelectron microscopy analysis (IEM) further provided evidence that EOB13250 was an endospore wall protein. These results together suggested that EOB13250 was a novel spore wall protein of N. bombycis. This study provides a further enrichment of the number of identified spore wall proteins in microsporidia and advances our understanding of the spore wall formation mechanism in these obligate unicellular parasites.
Substances chimiques
DNA, Protozoan
0
Protozoan Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
671-677Informations de copyright
© 2020 International Society of Protistologists.
Références
Bohne, W., Ferguson, D. J., Kohler, K. & Gross, U. 2000. Developmental expression of a tandemly repeated, glycine-and serine-rich spore wall protein in the microsporidian pathogen Encephalitozoon cuniculi. Infect. Immun., 68:2268-2275.
Briza, P., Breitenbach, M., Ellinger, A. & Segall, J. 1990. Isolation of two developmentally regulated genes involved in spore wall maturation in Saccharomyces cerevisiae. Genes Dev., 4:1775-1789.
Coluccio, A. E., Rodriguez, R. K., Kernan, M. J. & Neiman, A. M. 2008. The yeast spore wall enables spores to survive passage through the digestive tract of Drosophila. PLoS One, 3:e2873.
Dame, J. B., Williams, J. L., McCutchan, T. F., Weber, J. L., Wirtz, R. A., Hockmeyer, W. T., Maloy, W. L., Haynes, J. D., Schneider, I. & Roberts, D. 1984. Structure of the gene encoding the immunodominant surface antigen on the sporozoite of the human malaria parasite Plasmodium falciparum. Science, 225:593-599.
Ferguson, D. J., Balaban, A. E., Patzewitz, E.-M., Wall, R. J., Hopp, C. S., Poulin, B., Mohmmed, A., Malhotra, P., Coppi, A. & Sinnis, P. 2014. The repeat region of the circumsporozoite protein is critical for sporozoite formation and maturation in Plasmodium. PLoS One, 9:e113923.
Hayman, J. R., Hayes, S. F., Amon, J. & Nash, T. E. 2001. Developmental expression of two spore wall proteins during maturation of the microsporidian Encephalitozoon intestinalis. Infect. Immun., 69:7057-7066.
Hoyer, L. L., Green, C. B., Oh, S.-H. & Zhao, X. 2008. Discovering the secrets of the Candida albicans agglutinin-like sequence (ALS) gene family-a sticky pursuit. Med. Mycol., 46:1-15.
Jorda, J. & Kajava, A. V. 2009. T-REKS: identification of Tandem REpeats in sequences with a K-meanS based algorithm. Bioinformatics, 25:2632-2638.
Katinka, M. D., Duprat, S., Cornillot, E., Méténier, G., Thomarat, F., Prensier, G., Barbe, V., Peyretaillade, E., Brottier, P. & Wincker, P. 2001. Genome sequence and gene compaction of the eukaryote parasite Encephalitozoon cuniculi. Nature, 414:450.
Levdansky, E., Romano, J., Shadkchan, Y., Sharon, H., Verstrepen, K. J., Fink, G. R. & Osherov, N. 2007. Coding tandem repeats generate diversity in Aspergillus fumigatus genes. Eukaryot. Cell, 6:1380-1391.
Levdansky, E., Sharon, H. & Osherov, N. 2008. Coding fungal tandem repeats as generators of fungal diversity. Fungal Biol. Rev., 22:85-96.
Li, E., Yue, F., Chang, Q., Guo, X., He, X. & Zhang, B. 2013. Deletion of intragenic tandem repeats in unit C of FLO1 of Saccharomyces cerevisiae increases the conformational stability of flocculin under acidic and alkaline conditions. PLoS One, 8:e53428.
Li, Y., Tao, M., Ma, F., Pan, G., Zhou, Z. & Wu, Z. 2015. A monoclonal antibody that tracks endospore formation in the microsporidium Nosema bombycis. PLoS One, 10:e0121884.
Li, Z., Pan, G., Li, T., Huang, W., Chen, J., Geng, L., Yang, D., Wang, L. & Zhou, Z. 2012. SWP5, a spore wall protein, interacts with polar tube proteins in the parasitic microsporidian Nosema bombycis. Eukaryot. Cell, 11:229-237.
Luis Castillo, A.L.M., Garcera, A., Victoria Elorza, M., Valentin, E. & Sentandreu, R. 2003. Functional analysis of the cysteine residues and the repetitive sequence of Saccharomyces cerevisiae Pir4/Cis3: the repetitive sequence is needed for binding to the cell wall β-1,3-glucan. Yeast, 20:973-983.
Mendes, T., Lobo, F., Rodrigues, T., Rodrigues-Luiz, G., Darocha, W., Fujiwara, R., Teixeira, S. & Bartholomeu, D. 2013. Repeat-enriched proteins are related to host cell invasion and immune evasion in parasitic protozoa. Mol. Biol. Evol., 30:951-963.
Newman, A. M. & Cooper, J. B. 2007. XSTREAM: a practical algorithm for identification and architecture modeling of tandem repeats in protein sequences. BMC Bioinformat., 8:1.
Peuvel-Fanget, I., Polonais, V., Brosson, D., Texier, C., Kuhn, L., Peyret, P., Vivarès, C. & Delbac, F. 2006. EnP1 and EnP2, two proteins associated with the Encephalitozoon cuniculi endospore, the chitin-rich inner layer of the microsporidian spore wall. Int. J. Parasitol., 36:309-318.
Smits, G. J., van den Ende, H. & Klis, F. M. 2001. Differential regulation of cell wall biogenesis during growth and development in yeast. Microbiology, 147:781-794.
Southern, T. R., Jolly, C. E., Lester, M. E. & Hayman, J. R. 2007. EnP1, a microsporidian spore wall protein that enables spores to adhere to and infect host cells in vitro. Eukaryot. Cell, 6:1354-1362.
Tadakimi, T., Ma, Y. & Louis, W. 2018. Characterization of a SRS13: a new cyst wall mucin-like domain containing protein. Parasitol. Res, 117:2457-2466.
Van, T. T., Kim, S. K., Camps, M., Boothroyd, J. C. & Knoll, L. J. 2007. The BSR4 protein is up-regulated in Toxoplasma gondii bradyzoites, however the dominant surface antigen recognised by the P36 monoclonal antibody is SRS9. Int. J. Parasitol., 37:877-885.
Verstrepen, K. J., Jansen, A., Lewitter, F. & Fink, G. R. 2005. Intragenic tandem repeats generate functional variability. Nat. Genet., 37:986-990.
Wang, Y., Dang, X., Ma, Q., Liu, F., Pan, G., Li, T. & Zhou, Z. 2015. Characterization of a novel spore wall protein NbSWP16 with proline-rich tandem repeats from Nosema bombycis (microsporidia). Parasitology, 142:534-542.
Wang, Y., Geng, H., Dang, X., Xiang, H., Li, T., Pan, G. & Zhou, Z. 2017. Comparative analysis of the proteins with tandem repeats from 8 microsporidia and characterization of a novel endospore wall protein colocalizing with polar tube from Nosema bombycis. J. Eukaryot. Microbiol., 64:707-715.
Wesołowska, M., Szetela, B., Kicia, M., Kopacz, Ż., Sak, B., Rymer, W., Kváč, M. & Sałamatin, R. 2019. Dual infection of urinary tract with Enterocytozoon bieneusi and Encephalitozoon cuniculi in HIV/AIDS patients. Ann. Parasitol., 65:77-81.
Williams, B. A. & Keeling, P. J. 2011. 2 Microsporidia-highly reduced and derived relatives of fungi. Evolution of fungi and fungal-like organisms. Berlin, Heidelberg: Springer. p. 25-36.
Wittner, M. & Weiss, L. M. 1999. The microsporidia and microsporidiosis. ASM Press. Washington, DC.
Wu, Z., Li, Y., Pan, G., Tan, X., Hu, J., Zhou, Z. & Xiang, Z. 2008. Proteomic analysis of spore wall proteins and identification of two spore wall proteins from Nosema bombycis (microsporidia). Proteomics, 8:2447-2461.
Xu, Y., Takvorian, P., Cali, A., Wang, F., Zhang, H., Orr, G. & Weiss, L. M. 2006. Identification of a new spore wall protein from Encephalitozoon cuniculi. Infect. Immun., 74:239-247.
Zhao, W., Hao, Y., Wang, L., Zhou, Z. & Li, Z. 2015. Development of a strategy for the identification of surface proteins in the pathogenic microsporidian Nosema bombycis. Parasitology, 142:865-878.