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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: biohxh@fjnu.edu.cn
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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

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