caa a22 4500 445341521 CHVBK 20180317142814.0 cr unu---uuuuu 170323e20110801xx s 000 0 eng 10.1007/s00436-011-2270-2 doi (NATIONALLICENCE)springer-10.1007/s00436-011-2270-2 Different structure and mRNA expression of Entamoeba invadens chitinases in the encystation and excystation [Elektronische Daten] [Asao Makioka, Masahiro Kumagai, Kazushi Hiranuka, Seiki Kobayashi, Tsutomu Takeuchi] Entamoeba histolytica forms chitin-walled cysts during encystation process, where formation of the cyst wall needs not only chitin synthase but also chitinase. During excystation, quadruplet amoebae emerge from the chitin-walled cysts by dissolving the wall, so that chitinase may be necessary for excystation process as well. There is, however, no report on chitinase expression during excystation. In this study, we used Entamoeba invadens, a reptilian amoeba, as a model for encystation and excystation of E. histolytica, and studied chitinase mRNA expression in those processes. Although expression of three E. invadens chitinases designated EiChit1, EiChit2, and EiChit3 during encystation has been reported, we identified another enzyme named as EiChit4 in the E. invadens genome database. Therefore, we investigated the primary structure and mRNA expression of these four chitinases of Ei in the excystation as well as the encystation by real-time reverse transcription polymerase chain reaction (RT-PCR). Like EiChit1, EiChit4 had an 8 × Cys chitin-binding domain (CBD) and a hydrophilic spacer between the CBD and catalytic domain, and was also closer to EiChit1 than EiChit2 and EiChit3 in the phylogenetic tree. During encystation, the expression of all four chitinases increased in the early phase; the increase in EiChit1 and EiChit4 was much higher than in EiChit2 and EiChit3. Then, the expression of all four chitinases sharply decreased in the later phase. In cysts, EiChit1 was most abundantly expressed and EiChit4 was at a lower level, while the expressions of EiChit2 and EiChit3 were virtually absent. Following the induction of excystation, mRNA levels of EiChit1 and EiChit4 in cysts 5h after induction were significantly lower than those in cysts before induction, while those of EiChit2 and EiChit3 were remarkably higher than before induction. The mRNAs of only EiChit2 and EiChit3 remarkably increased when the excystation was induced in the presence of cytochalasin D. These data demonstrate different structures and expressions of four chitinases in the differentiation of E. invadens. Springer-Verlag, 2011 Makioka Asao Department of Tropical Medicine, Jikei University School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, 105-8461, Tokyo, Japan aut Kumagai Masahiro Department of Tropical Medicine, Jikei University School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, 105-8461, Tokyo, Japan aut Hiranuka Kazushi Institute for Chemical Research, Kyoto University, Gokasho, Uji, 611-0011, Kyoto, Japan aut Kobayashi Seiki Department of Tropical Medicine and Parasitology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, 160-8582, Tokyo, Japan aut Takeuchi Tsutomu Department of Tropical Medicine and Parasitology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, 160-8582, Tokyo, Japan aut Parasitology Research Springer-Verlag 109/2(2011-08-01), 417-423 0932-0113 109:2<417 2011 109 436 https://doi.org/10.1007/s00436-011-2270-2 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00436-011-2270-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Makioka Asao Department of Tropical Medicine, Jikei University School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, 105-8461, Tokyo, Japan aut NATIONALLICENCE 700 1- Kumagai Masahiro Department of Tropical Medicine, Jikei University School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, 105-8461, Tokyo, Japan aut NATIONALLICENCE 700 1- Hiranuka Kazushi Institute for Chemical Research, Kyoto University, Gokasho, Uji, 611-0011, Kyoto, Japan aut NATIONALLICENCE 700 1- Kobayashi Seiki Department of Tropical Medicine and Parasitology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, 160-8582, Tokyo, Japan aut NATIONALLICENCE 700 1- Takeuchi Tsutomu Department of Tropical Medicine and Parasitology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, 160-8582, Tokyo, Japan aut NATIONALLICENCE 773 0- Parasitology Research Springer-Verlag 109/2(2011-08-01), 417-423 0932-0113 109:2<417 2011 109 436 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer