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Molecular characterization and expression analysis of three novel autophagy-related genes from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

Published online by Cambridge University Press:  09 September 2016

JOSÉ MIGUEL FLORES FERNÁNDEZ ,
CARLA PATRICIA BARRAGÁN ÁLVAREZ ,
CARLA VANESSA SÁNCHEZ HERNÁNDEZ ,
EDUARDO PADILLA CAMBEROS ,
CELIA GONZÁLEZ CASTILLO ,
DANIEL ORTUÑO SAHAGÚN  and
MOISÉS MARTÍNEZ VELÁZQUEZ
JOSÉ MIGUEL FLORES FERNÁNDEZ
Affiliation:
Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, AC, Av. Normalistas 800, Col. Colinas de la Normal, 44270. Guadalajara, Jalisco, México
CARLA PATRICIA BARRAGÁN ÁLVAREZ
Affiliation:
Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, AC, Av. Normalistas 800, Col. Colinas de la Normal, 44270. Guadalajara, Jalisco, México
CARLA VANESSA SÁNCHEZ HERNÁNDEZ
Affiliation:
Departamento de Producción Agrícola, CUCBA, Universidad de Guadalajara, Carretera Guadalajara-Nogales km 15.5, 45110, Zapopan, Jalisco, México
EDUARDO PADILLA CAMBEROS
Affiliation:
Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, AC, Av. Normalistas 800, Col. Colinas de la Normal, 44270. Guadalajara, Jalisco, México
CELIA GONZÁLEZ CASTILLO
Affiliation:
Instituto de Investigación en Ciencias Biomédicas (IICB), CUCS, Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340. Guadalajara, Jalisco, México
DANIEL ORTUÑO SAHAGÚN
Affiliation:
Instituto de Investigación en Ciencias Biomédicas (IICB), CUCS, Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340. Guadalajara, Jalisco, México
MOISÉS MARTÍNEZ VELÁZQUEZ*
Affiliation:
Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, AC, Av. Normalistas 800, Col. Colinas de la Normal, 44270. Guadalajara, Jalisco, México
*
*Corresponding author: Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, AC, Av. Normalistas 800, Col. Colinas de la Normal, 44270. Guadalajara, Jalisco, México. E-mail: [email protected]
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Summary

The cattle tick Rhipicephalus (Boophilus) microplus is a hematophagous ectoparasite of major importance for the livestock industry. It shows a remarkable ability to survive over long periods without feeding. However, the mechanisms used to endure long-term starvation are poorly understood. It is believed that autophagy, a process of intracellular protein degradation, may play a significant role to confront adverse environmental conditions. To advance our understanding of autophagy in R. microplus, in the present study we report the molecular characterization of three autophagy-related (ATG) genes, namely, RmATG3, RmATG4 and RmATG6, as well as their expression profiles in different developmental stages and organs of the parasite. The deduced amino acid sequences derived from the characterized gene sequences were subjected to Basic Local Alignment Search Tool analysis. The testing produced significant alignments with respective ATG proteins from Haemaphysalis longicornis and Ixodes scapularis ticks. Real-time polymerase chain reaction assays revealed that RmATG4 and RmATG6 transcripts were elevated in egg and ovary tissue, when compared with larva and midgut samples, while RmATG3 expression in midgut was 2-fold higher than in egg, larva and ovary samples.

Keywords

AutophagyATG3ATG4ATG6Rhipicephalus microplusreal-time PCRdevelopmentembryogenesis
Type
Research Article
Information
Parasitology , Volume 143 , Issue 13 , November 2016 , pp. 1802 - 1809
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Copyright
Copyright © Cambridge University Press 2016 

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References

REFERENCES

Altschul, S. F., Gish, W., Miller, W., Myers, E. W. and Lipman, D. J. (1990). Basic local alignment search tool. Journal of Molecular Biology 215, 403–410.Google Scholar
Flores Fernández, J. M., Gutiérrez Ortega, A., Rosario Cruz, R., Padilla Camberos, E., Alvarez, A. H. and Martínez Velázquez, M. (2014). Molecular cloning and characterization of two novel autophagy-related genes belonging to the ATG8 family from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae). Experimental and Applied Acarology 64, 533–542.Google Scholar
Gasteiger, E., Gattiker, A., Hoogland, C., Ivanyi, I., Appel, R. D. and Bairoch, A. (2003). ExPASy: the proteomics server for in-depth protein knowledge and analysis. Nucleic Acids Research 31, 3784–3788.Google Scholar
Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, M. R., Appel, R. D. and Bairoch, A. (2005). Protein identification and analysis tools on the ExPASy server. In The proteomics Protocols Handbook (ed. Walker John, M.), pp. 571–607. Humana Press, New York.Google Scholar
Kawano, S., Umemiya-Shirafuji, R., Boldbaatar, D., Matsuoka, K., Tanaka, T. and Fujisaki, K. (2011). Cloning and characterization of the autophagy-related gene 6 from the hard tick, Haemaphysalis longicornis . Parasitology Research 109, 1341–1349.CrossRefGoogle ScholarPubMed
Larkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., McGettigan, P. A., McWilliam, H., Valentin, F., Wallace, I. M., Wilm, A., Lopez, R., Thompson, J. D., Gibson, T. J. and Higgins, D. G. (2007). Clustal W and Clustal X version 2.0. Bioinformatics 23, 2947–2948.Google Scholar
Livak, K. J. and Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25, 402–408.Google Scholar
McPhee, C. K. and Baehrecke, E. H. (2009). Autophagy in Drosophila melanogaster . Biochimica et Biophysica Acta 1793, 1452–1460.Google Scholar
Mizushima, N. and Sahani, M. H. (2014). ATG8 localization in apicomplexan parasites: apicoplast and more. Autophagy 10, 1487–1494.Google Scholar
Mizushima, N., Yoshimori, T. and Levine, B. (2010). Methods in mammalian autophagy research. Cell 140, 313–326.Google Scholar
Nijhof, A. M., Balk, J. A., Postigo, M. and Jongejan, F. (2009). Selection of reference genes for quantitative RT-PCR studies in Rhipicephalus (Boophilus) microplus and Rhipicephalus appendiculatus ticks and determination of the expression profile of Bm86. BMC Molecular Biology 10, 112.Google Scholar
Rozen, S. and Skaletsky, H. J. (2000). Primer3 on the WWW for general users and for biologist programmers. In Bioinformatics Methods and Protocols: Methods in Molecular Biology (ed. Krawetz, S. and Misener, S.), pp. 365–386. Humana, Totowa.Google Scholar
Sharma, A. K., Kumar, R., Kumar, S., Nagar, G., Singh, N. K., Rawat, S. S., Dhakad, M. L., Rawat, A. K., Ray, D. D. and Ghosh, S. (2012). Deltamethrin and cypermethrin resistance status of Rhipicephalus (Boophilus) microplus collected from six agro-climatic regions of India. Veterinary Parasitology 188, 337–345.CrossRefGoogle ScholarPubMed
Spickler, A. R. (2007). Rhipicephalus (Boophilus) Microplus. http://www.cfsph.iastate.edu/DiseaseInfo/factsheets.php.Google Scholar
Tsuda, A., Mulenga, A., Sugimoto, C., Nakajima, M., Ohashi, K. and Onuma, M. (2001). cDNA cloning, characterization and vaccine effect analysis of Haemaphysalis longicornis tick saliva proteins. Vaccine 19, 4287–4296.Google Scholar
Umemiya-Shirafuji, R., Matsuo, T., Liao, M., Boldbaatar, D., Battur, B., Suzuki, H. and Fujisaki, K. (2010). Increased expression of ATG genes during nonfeeding periods in the tick Haemaphysalis longicornis . Autophagy 6, 473–481.Google Scholar
Umemiya-Shirafuji, R., Galay, R. L., Maeda, H., Kawano, S., Tanaka, T., Fukumoto, S., Suzuki, H., Tsuji, N. Fujisaki, K. (2014). Expression analysis of autophagy-related genes in the hard tick Haemaphysalis longicornis . Veterinary Parasitology 201, 169–175.Google Scholar
Xie, Z. and Klionsky, D. J. (2007). Autophagosome formation: core machinery and adaptations. Nature Cell Biology 9, 1102–1109.Google Scholar
Zhang, X., Hu, Z. Y., Li, W. F., Li, Q. R., Deng, X. J., Yang, W. Y., Cao, Y. and Zhou, C. Z. (2009). Systematic cloning and analysis of autophagy-related genes from the silkworm Bombyx mori . BMC Molecular Biology 10, 50.CrossRefGoogle ScholarPubMed
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