Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-25T02:03:01.635Z Has data issue: false hasContentIssue false
  • English
  • Français

Molecular characterization of an actin depolymerizing factor from Cryptocaryon irritans

Published online by Cambridge University Press:  04 January 2013

XIAOHONG HUANG ,
YANG XU ,
GUOWEI GUO ,
QIANQIAN LIN ,
ZHONGFENG YE ,
LIPING YUAN ,
ZHIYU SUN  and
WEI NI
XIAOHONG HUANG*
Affiliation:
Fujian Provincial Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350108, Fujian, China
YANG XU
Affiliation:
Fujian Provincial Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350108, Fujian, China
GUOWEI GUO
Affiliation:
Fujian Provincial Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350108, Fujian, China
QIANQIAN LIN
Affiliation:
Fujian Provincial Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350108, Fujian, China
ZHONGFENG YE
Affiliation:
Fujian Provincial Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350108, Fujian, China
LIPING YUAN
Affiliation:
Fujian Provincial Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350108, Fujian, China
ZHIYU SUN
Affiliation:
Fujian Provincial Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350108, Fujian, China
WEI NI
Affiliation:
Fujian Provincial Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350108, Fujian, China
*
*Corresponding author: Fujian Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Qishan Campus, Fuzhou 350108, Fujian, China. Tel: +86 15859107668. Fax: +86 0591 22868199. E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

Actin depolymerizing factors regulate actin dynamics involved in cellular processes such as morphogenesis, motility, development and infection. Here, a novel actin depolymerizing factor gene (CiADF2) was cloned from the cDNA library of Cryptocaryon irritans, a parasitic ciliate causing cryptocaryonosis. The full-length cDNA of CiADF2 was 531 bp. Its open reading frame (ORF) was 417 bp, encoding a polypeptide of 138 aa with typical features of the ADF/cofilin family. Reverse transcription-PCR suggested that CiADF2 is expressed in all stages of the life cycle. After site-directed mutagenesis of a non-universal genetic code, the ORF was subcloned in Escherichia coli. The bacteria were induced with the addition of isopropylthio-β-D-galactoside to express a fusion protein of recombinant CiADF2 (rCiADF2) with glutathione S transferase. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot confirmed the predicted molecular mass of rCiADF2 of 16·2 kDa. A mouse antibody against rCiADF2 recognized native CiADF2, and rCiADF2 reacted with mouse antisera against C. irritans trophonts. CiADF2 was abundant in the plasma around cytostomes, suggesting that CiADF2 is involved in ciliate movement. Moreover, rCiADF2 showed F-actin binding and depolymerizing activity. This study will help to clarify the pathogenic biology of the parasite and develop effective control measures for cryptocaryonosis.

Keywords

Cryptocaryon irritansactin depolymerizing factormolecular characterization
Type
Research Article
Information
Parasitology , Volume 140 , Issue 5 , April 2013 , pp. 561 - 568
Copyright
Copyright © Cambridge University Press 2013

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

REFERENCES

Allen, M. L., Dobrowolski, J. M., Muller, H., Sibley, L. D. and Mansour, T. E. (1997). Cloning and characterization of actin depolymerising factor from Toxoplasma gondii . Molecular and Biochemical Parasitology 88, 43–52.CrossRefGoogle ScholarPubMed
Bernstein, B. W. and Bamburg, J. R. (2010). ADF/cofilin: a functional node in cell biology. Trends in Cell Biology 20, 187–195.CrossRefGoogle ScholarPubMed
Colorni, A. and Burgess, P. (1997). Cryptocaryon irritans Brown 1951, the cause of white spot disease in marine fish: an update. Aquarium Sciences and Conservation 1, 217–238.Google Scholar
Diggles, B. K. and Adlard, R. D. (1997). Intraspecific variation in Cryptocaryon irritans . The Journal of Eukaryotic Microbiology 44, 25–32.CrossRefGoogle ScholarPubMed
Doi, Y., Shinzawa, N., Fukumoto, S., Okano, H. and Kanuka, H. (2010). ADF2 is required for transformation of the ookinete and sporozoite in malaria parasite development. Biochemical and Biophysical Research Communications 397, 668–672.Google Scholar
Huang, X., Sun, Z., Guo, G., Zheng, C., Xu, Y., Yuan, L. and Liu, C. (2012). Cloning and characterization of a surface antigen CiSA-32·6 from Cryptocaryon irritans . Experimental Parasitology 130, 189–194.Google Scholar
Huang, X., Xuan, X., Yokoyama, N., Xu, L., Suzuki, H., Sugimoto, C., Nagasawa, H., Fujisaki, K. and Igarashi, I. (2003). High-level expression and purification of a truncated merozoite antigen-2 of Babesia equi in Escherichia coli and its potential for immunodiagnosis. Journal of Clinical Microbiology 41, 1147–1151.Google Scholar
Kumar, G., Srivastava, R., Mitra, K., Sahasrabuddhe, A. A. and Gupta, C. M. (2012). Overexpression of S4D mutant of Leishmania donovani ADF/Cofilin impairs flagellum assembly by affecting actin dynamics. Eukaryotic Cell 11, 752–760.Google Scholar
Makioka, A., Kumagai, M., Hiranuka, K., Kobayashi, S. and Takeuchi, T. (2011). Entamoeba invadens: identification of ADF/cofilin and their expression analysis in relation to encystation and excystation. Experimental Parasitology 127, 195–201.CrossRefGoogle ScholarPubMed
Mehta, S., and Sibley, L. D. (2011). Actin depolymerizing factor controls actin turnover and gliding motility in Toxoplasma gondii . Molecular Biology of the Cell 22, 1290–1299.CrossRefGoogle ScholarPubMed
Pollard, T. D., and Borisy, G. G. (2003). Cellular motility driven by assembly and disassembly of actin filaments. Cell 112, 453–465.CrossRefGoogle ScholarPubMed
Sibley, L. D. (2004). Intracellular parasite invasion strategies. Science 304, 248–253.CrossRefGoogle ScholarPubMed
Sun, H. Y., Zhu, X. Q., Xie, M. Q., Wu, X. Y., Li, A. X., Lin, R. Q. and Song, H. Q. (2006). Characterization of Cryptocaryon irritans isolates from marine fishes in Mainland China by ITS ribosomal DNA sequences. Parasitology Research 99, 160–166.CrossRefGoogle ScholarPubMed
Sun, Z., Zheng, C., Wu, X., Guo, G., Wang, Y. and Huang, X. (2011). The strain and life-cycle of Cryptocaryon irritans isolated from Pseudosciaena crocea cultured in Xiapu, Fujian. Journal of Fujian Normal University (Natural Science Edition) 27, 101–108.Google Scholar
Tammana, T. V., Sahasrabuddhe, A. A., Bajpai, V. K., Gupta, C. M. (2010). ADF/cofilin-driven actin dynamics in early events of Leishmania cell division. Journal of Cell Science 123, 1894–1901.Google Scholar
Tammana, T. V., Sahasrabuddhe, A. A., Mitra, K., Bajpai, V. K. and Gupta, C. M. (2008). Actin-depolymerising factor, ADF/cofilin, is essentially required in assembly of Leishmania flagellum . Molecular Microbiology 70, 837–852.CrossRefGoogle Scholar
Xu, J. H., Qin, Z. H., Liao, Y. S., Xie, M. Q., Li, A. X. and Cai, J. P. (2008). Characterization and expression of an actin-depolymerising factor from Eimeria tenella . Parasitology Research 103, 263–270.Google Scholar
Zhou, B. H., Wang, H. W., Xue, F. Q., Wang, X. Y., Yang, F. K., Ban, M. M., Xin, R. X. and Wang, C. C. (2010). Actin-depolymerising factor of second-generation merozoite in Eimeria tenella: clone, prokaryotic expression, and diclazuril-induced mRNA expression. Parasitology Research 106, 571–576.Google Scholar