폐흡충은 국내를 비롯한 아시아에서 폐흡충증을 일으키는 중요한 기생충이다. 이러한 폐흡충은 이배체 와 삼배체가 국내에 알려져 있다. 이배체 폐흡충은 양성생식을 하고 삼배체 폐흡충은 단위생식을 하는 것으로 알려져 있다. 그러나 이러한 원인에 대하여서는 알려진 바가 없다. 생식유전자 중 초파리에서 최초 분리되었고 포유동물에서도 그 기능이 밝혀진 vasa 유전자가 가장 유명하다. 이 유전자는 생식세포의 분화에 관여하며 종에 따라서는 정자생성에도 관여하는 것으로 알려져 있다. 이번 연구는 삼배체 폐흡충의 단위생식과 vasa 유전자와의 관계를 규명해 보고자 하였다. 폐흡충의 이배체와 삼배체 성충으로부터 vasa 유전자 전체의 염기서열을 얻을 수 있었다. 두 염기서열의 경우 8개의 ATP-binding domain이 관찰되었고 helicase가 결합할 것으로 예상되는 RGG motif도 관찰되었다. 총 622아미노산 서열로 구성될 것으로 보이며 이배체는 69.018 kDa, 삼배체는 68.930 kDa 크기의 단백질을 만들 것으로 예상되었다. Vasa 재조합 단백질은 GST와 fusion되어 93 kDa 크기에서 관찰되었다. mRNA의 발현은 이배체에 비해 삼배체가 다소 높았다. Anti-Pw-VASA항체를 이용한 면역조직화학법을 수행한 결과 이배체와 삼배체는 다른 기관에서는 면역반응력을 보이지 않고 고환에서만 면역반응력을 볼 수 있었다. 결과를 종합해 보면 vasa 유전자는 이배체 삼배체 모두 가지고 있었으며 정상적으로 발현되었다.
폐흡충은 국내를 비롯한 아시아에서 폐흡충증을 일으키는 중요한 기생충이다. 이러한 폐흡충은 이배체 와 삼배체가 국내에 알려져 있다. 이배체 폐흡충은 양성생식을 하고 삼배체 폐흡충은 단위생식을 하는 것으로 알려져 있다. 그러나 이러한 원인에 대하여서는 알려진 바가 없다. 생식유전자 중 초파리에서 최초 분리되었고 포유동물에서도 그 기능이 밝혀진 vasa 유전자가 가장 유명하다. 이 유전자는 생식세포의 분화에 관여하며 종에 따라서는 정자생성에도 관여하는 것으로 알려져 있다. 이번 연구는 삼배체 폐흡충의 단위생식과 vasa 유전자와의 관계를 규명해 보고자 하였다. 폐흡충의 이배체와 삼배체 성충으로부터 vasa 유전자 전체의 염기서열을 얻을 수 있었다. 두 염기서열의 경우 8개의 ATP-binding domain이 관찰되었고 helicase가 결합할 것으로 예상되는 RGG motif도 관찰되었다. 총 622아미노산 서열로 구성될 것으로 보이며 이배체는 69.018 kDa, 삼배체는 68.930 kDa 크기의 단백질을 만들 것으로 예상되었다. Vasa 재조합 단백질은 GST와 fusion되어 93 kDa 크기에서 관찰되었다. mRNA의 발현은 이배체에 비해 삼배체가 다소 높았다. Anti-Pw-VASA항체를 이용한 면역조직화학법을 수행한 결과 이배체와 삼배체는 다른 기관에서는 면역반응력을 보이지 않고 고환에서만 면역반응력을 볼 수 있었다. 결과를 종합해 보면 vasa 유전자는 이배체 삼배체 모두 가지고 있었으며 정상적으로 발현되었다.
In this study, we isolated, characterized, and compared the vasa homologous genes of diploid and triploid Paragonimus westermani and localized VASA homologous proteins in both lung fluke types. Open reading frames of Pw-vasa-2n and Pw-vasa-3n were of 1812 bp, and encoded deduced proteins of 622 amin...
In this study, we isolated, characterized, and compared the vasa homologous genes of diploid and triploid Paragonimus westermani and localized VASA homologous proteins in both lung fluke types. Open reading frames of Pw-vasa-2n and Pw-vasa-3n were of 1812 bp, and encoded deduced proteins of 622 amino acids with calculated molecular weights of 69.0 kDa and 68.9 kDa and pI's of 9.11 and 9.03, respectively. A comparison of these two VASA deduced protein sequences showed that only 6 of the 622 amino acids differed. The deduced sequences of Pw-VASA-2n and Pw-VASA-3n contained eight consensus sequences characteristic of the DEAD-box protein family and their N-terminal regions contained four arginine-glycine-glycine (RGG) motifs. These two lung fluke VASA-like proteins were more similar to those of other VASA proteins than to those of other DEAD-family proteins isolated from several organisms (planarian, zebra fish, mouse, and human). vasa homologous gene transcription and VASA protein expressions in triploid type lung flukes was slightly stronger than in the diploid type. Immunostaining showed that testes and a portion of the ovaries of both diploid and triploid lung flukes reacted strongly to anti-Pw-VASA antibody.
In this study, we isolated, characterized, and compared the vasa homologous genes of diploid and triploid Paragonimus westermani and localized VASA homologous proteins in both lung fluke types. Open reading frames of Pw-vasa-2n and Pw-vasa-3n were of 1812 bp, and encoded deduced proteins of 622 amino acids with calculated molecular weights of 69.0 kDa and 68.9 kDa and pI's of 9.11 and 9.03, respectively. A comparison of these two VASA deduced protein sequences showed that only 6 of the 622 amino acids differed. The deduced sequences of Pw-VASA-2n and Pw-VASA-3n contained eight consensus sequences characteristic of the DEAD-box protein family and their N-terminal regions contained four arginine-glycine-glycine (RGG) motifs. These two lung fluke VASA-like proteins were more similar to those of other VASA proteins than to those of other DEAD-family proteins isolated from several organisms (planarian, zebra fish, mouse, and human). vasa homologous gene transcription and VASA protein expressions in triploid type lung flukes was slightly stronger than in the diploid type. Immunostaining showed that testes and a portion of the ovaries of both diploid and triploid lung flukes reacted strongly to anti-Pw-VASA antibody.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
제안 방법
In this study, we isolated, characterized, and compared the vasa homologous gene of diploid and triploid Paragonimus westermani and localized VASA homologous proteins in both lung flukes. The predicted amino acid sequences encoded by these genes shared eight well-conserved domains of DEAD box family proteins, suggesting that they are ATP-dependent RNA helicasez including ATP~bindingz RNA-binding, and in-winding modalities [19, 20].
성능/효과
In conclusion, in the present study we isolated vasa genes for the first time from the diploid and triploid types of P. westermani. Our observations suggest that they function in the reproductive system, much as has been reported in other organisms.
westermani is aberrant and normal spermatozoa are not produced [6]. Moreover, their results show that P. pulmonalis (P. westermani triploid type) spermatogenesis is almost completely inhibited by aberrations in the process. The spermatozoa found in the testis of triploid type lung fluke may degenerate, because no spermatozoa were observed in seminal receptacles.
These results show that the two isolated VASA-like proteins have similar or identical functions. The identity rates of these two proteins was found to be highest with VASA proteins in planarian (61% and 57% similarity with diploid and triploid VASA proteins, respectively), in zebra fish (52%z 47%), in mouse (48%, 48%), and in man (48%, 48%), These similarities were substantially higher than those of other DEAD-family proteins, e.g., p68 in man (39%z 39%) or elF4A in mouse (32%, 32%, respectively). An analysis of the phylogenic relationships between members of the DEAD-box protein family showed three distinct clusters: the VASA sub-family (including the predicted lung flukes VASA sequences), the 이F4A sub-family, and p68 sub-family (Fig.
also examined the chromosome karyotype, and some genes of two flukes isolated in Haenam and Youngam provinces (Korea) [17, 18]. The results obtained showed that lung flukes isolated in Haenam province were diploid, while those from Youngam province were triploid. These results supported the hypothesis that diploid lung flukes have functional sexual organs and normally reproduce sperm and eggs, whereas triploid lung fluke cannot reproduce sperm because reproductive organs do not function [6].
참고문헌 (25)
Blair, D., T. Agatsuma, T. Watanabe, M. Okamoto and A. Ito. 1997. Geographical genetic structure within the human lung fluke, Paragonimus westermani, detected from DNA sequences. Parasitol. 115, 411-417
Blair, D., Z. B. Xu. and T. Agatsuma. 1999. Paragonimiasis and the Genus Paragonimus. Adv Parasit 42, 113-222
Castril1on, D. H., B. J. Quade, T. Y. Wang, C. Quigley and C. P. Crum. 2000. The human VASA gene is specifically expressed in the germ cell lineage. Proc. Natl. Acad. Sci. U.S.A. 97, 9585-9590
Eres, R. 2000. The fuction and regulation of vasa-like genes in germ cell development. Genom. Bio.l 1, 1017.1-1017.6
Fan, P. C. and C. H. Chiang. 1970. Exposure of kittens and puppies to single metacercariae of Paragonimus westermani from Taiwan. J. Parasitol. 56, 48-54
Fujino, T. and Y. Ishii. 1982. Ultrastructural studies on spermatogenesis in a parthenogenetic type of Paragonimus westermani (Kerbert 1978) proposed as P. pulminalis (Baelz 1880). J. Parasitol. 68, 433-441
Fujiwara, Y., T. Komiya, H. Kawabata, M. Sato, H. Fujimoto, M. Furusawa and T. Noce. 1994. Isolation of a DEAD-family protein gene that encodes a murine homolog of Drosophila vasa and its specific expression in germ cell lineage. Proc. Natl. Acad. Sci. U.S.A. 91, 12258-12262
Hay, B., L. Y. Jan and Y. N. Jan. 1988. A protein component of Drosophila polar granules is encoded by vasa and has extensive sequence similarity to ATP-dependent helicases. Cell 55, 577-587
Ihm, G. S. and Y. K. Ahn 1979. Comparative studies on type determinant of Paragonimus westermami in Korea. Yonsei Med. J. 12, 38-47
Kobayashi S. and R. Amikura. 1993. Okada M. Presence of mitochondrial large ribosomal RNA outside mitochondria in germ plasm of Drosophila melanogaster. Science 260, 1521-1524
Liang, L., W. Diehl-Jones and P. Lasko. 1994. Localization of vasa protein to the Drosophila pole plasm is independent of its RNA-binding and helicase activities. Development 120, 1201-1211
Linder, P. 2000. Quick guide-DEAD-box proteins. Curr. Biol. 10, 887
Marchler-Bauer, A., J. B. Anderson, C. DeWeese-Scott, N. D. Fedorova, L. Y. Geer, He S, D. I Hurwitz, J. D. Jackson, A.R. Jacobs, C. J. Lanczycki, C. A. Liebert, C. Liu, T. Madej, G. H. Marchler, R. Mazumder, A. N. Nikolskaya, A. R. Panchenko, B. S. Rao, B. A. Shoemaker, V. Simonyan, J. S. Song, P. A .Thiessen, S. Vasudevan, Y. Wang, R. A.Yamashita, J. J .Yin and S. H. Bryant. 2003. CDD: a curated Entrez database of conserved domain alignments. Nucleic Acids Res. 31, 383-387
Miyazaki, I. 1978. Two types if lung fluke which has been called, Paragonimus westermani (Kerbert 1878). Medical Bulletin of Fukuoka University 5, 251-263
Miyazaki, I. 1982. Paragonimiasis. In Handbook Series in Zoonoses, Section C. Parasitic zoonoses, Hilyer GV and Hopla CE (eds.). CRC Press Inc, Boca Raton, USA, pp. 143-164
Park, G. M., K. I. Im and T. S. Yang. 2003. Phylogenetic relationship of ribosomal ITS2 and mitochondrial COI among diploid and triploid Paragonimus westermani isolates. Korean J. Parasitol. 41, 47-55
Park, G. M., K. J. Lee, K. I. Im, H. Park and T. S. Yang. 2001. Occurrence of a diploid type and a new first intermediate host of a human lung fluke, Paragonimus westermani, in Korea. Exp. Parasitol. 99, 206-212
Pause, A. and N. Sonenberg. 1992. Mutational analysis of a DEAD box RNA helicase: The mammalian translation initiation factor eIF-4A. EMBO J. 11, 2643-2654
Pause, A., N. Melhot and N. Sonenberg. 1993. The HRIGRXXR region of the DEAD box RNA helicase eukaryotic translation initiation factor 4A is required for RNA binding and ATP hydrolysis. Mol. Cell. Biol. 13, 6789-6798
Shibata, N., Y. Umesono, H. Orii, T. Sakurai, K. Watanabe and K. Agata. 1999. Expression of vasa (vas)-related genes in germline cells and totipotent somatic stem cells of planarians. Dev. Biol. 206, 73-87
Strome, S. 1992. Developmental biology. The germ of the issue. Nature (London) 358, 368-369
Tanaka, S. S., Y. Toyooka, R. Akasu, Y. Katoh-Fukui, Y. Nakahara, R. Suzuki, M. Yokoyama and T. Noce. 2000. The mouse homolog of Drosophila Vasa is required for the development of male germ cells. Genes Dev. 14, 841-853
Yokogawa, M. 1965. Paragonimus and paragonimiasis. Adv. Parasitol. 3, 99-158
Yokogawa, M., K. Araki, H. Koyama, B. S. Sea, S. H. Lee and S. Y. Cho. 1940. On the lung fluke, Paragonimus iloktsuenensis Chen, in Korea. Japanese J. Parasitol. 20, 215-221
※ AI-Helper는 부적절한 답변을 할 수 있습니다.