ISSN : 1225-3480
동양달팽이의 arginine kinase 유전자는 염기서열 1065개로 이루어져있으며 355개의 아미노산으로 이루어져 있으며,BLAST 결과를 토대로 유사도가 높은 25개의 참고 서열과 동양달팽이의 arginine kinase의 아미노산 서열을 MEGA5 프로그램의 clustalW 모듈을 이용하여 multiple sequencealignment 를 수행한 결과, 연체동물문에 속하는 복족강 (5종), 두족강 (5종), 이매패강 (4종) 등에 속하는 생물들이 같은군으로 묶였으며, 절지동물문 곤충강 에 속하는 나비목 (2종),벌목 (1종), 노린재목 (2종) 등에 속하는 생물들이 같은 군으로묶이고, 갑각강 (5종), 거미강 (1종) 에 속하는 생물들이 마지막으로 묶이는 것을 알 수 있었다. Psipred 소프트웨어를 통해 2D 구조를 비교 분석 한 결과도 multiple align 및phylodendrogram 결과와 밀접한 관계가 있음을 알 수 있었다. 시간에 따른 arginine kinase의 발현양상을 확인한 결과control에 비하여 6시간에서 약 1.2배 정도 발현이 증가 되는것을 확인할 수 있었으며, 12시간이 지나면 점차 감소하는 것을 확인할 수 있었다. EST를 통해 밝혀진 N. samarangae 의 Ark 서열은 근연종들의 서열과 일치함을 알 수 있었으며,본 연구 결과를 통해 무척추동물에서의 선천성 면역 관련 유전자 연구에 동양달팽이가 좋은 모델이 될 수 있음을 제시하고있다.
Arginine kinase (ArK) is known to play an important role in maintaining the level of ATP by phosphorylation of phosphagens in cell and immuninty in most living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.
Adams, R.J., Ewing, J., Dujovny, M., and Misra, M. (1998) Editorial commentary. J. Stroke Cerebrovasc Dis., 7: I-IV.
Bettencourt, R., Dando, P., Collins, P., Costa, V., Allam, B., and Serrao Santos, R. (2009) Innate immunity in the deep sea hydrothermal vent mussel Bathymodiolus azoricus. Comp. Biochem. Physiol. A. Mol. Integr. Physiol., 152: 278-289.
Biswas, C., and Mandal, C. (1999) The role of amoebocytes in endotoxin-mediated coagulation in the innate immunity of Achatina fulica snails. Scand. J. Immunol., 49: 131-138.
Bragg, J., Rajkovic, A., Anderson, C., Curtis, R., Van Houten, J., Begres, B., Naples, C., Snider, M., Fraga, D., and Singer, M. (2012) Identification and characterization of a putative arginine kinase homolog from Myxococcus xanthus required for fruiting body formation and cell differentiation. J. Bacteriol., 194: 2668-2676.
Braun, R.C., Pedretti, K.T., Casavant, T.L., Scheetz, T.E., Birkett, C.L., and Roberts, C.A. (2001) Parallelization of local BLAST service on workstation clusters. Future Generation Computer Systems, 17: 745-754.
Buermans, H.P., Ariyurek, Y., van Ommen, G., den Dunnen, J.T., and t Hoen, P.A. (2010) New methods for next generation sequencing based microRNA expression profiling. BMC Genomics, 11: 716.
Burge, C., and Karlin, S. (1997) Prediction of complete gene structures in human genomic DNA. J. Mol. Biol., 268: 78-94.
Burge, C.B., and Karlin, S. (1998) Finding the genes in genomic DNA. Curr. Opin. Struct. Biol., 8: 346-354.
Charlet, M., Chernysh, S., Philippe, H., Hetru, C., Hoffmann, J.A., and Bulet, P. (1996) Innate immunity. Isolation of several cysteine-rich antimicrobial peptides from the blood of a mollusc, Mytilus edulis. J. Biol. Chem., 271: 21808-21813.
Chistoserdovai, L. (2010) Functional metagenomics: recent advances and future challenges. Biotechnol. Genet. Eng. Rev., 26: 335-352.
Coyne, V. (2011) The importance of ATP in the immune system of molluscs. Invertebrate Survival Journal, 8: 48-55.
Cui, S., Zhang, D., Jiang, S., Pu, H., Hu, Y., Guo, H., Chen, M., Su, T., and Zhu, C. (2011) A macrophage migration inhibitory factor like oxidoreductase from pearl oyster Pinctada fucata involved in innate immune responses. Fish Shellfish Immunol, 31: 173-181.
Dumas, C., and Camonis, J. (1993) Cloning and sequence analysis of the cDNA for arginine kinase of lobster muscle. J. Biol. Chem., 268: 21599-21605.
Edgar, R.C. (2004) MUSCLE: a multiple sequence alignment method with reduced time and space complexity. BMC Bioinformatics, 5: 113.
Edgar, R.C. (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res., 32: 1792-1797.
Ewing, B., and Green, P. (1998) Base-calling of automated sequencer traces using phred. II. Error probabilities. Genome Res., 8: 186-194.
Ewing, B., Hillier, L., Wendl, M.C., and Green, P. (1998) Base-calling of automated sequencer traces using phred. I. Accuracy assessment. Genome Res., 8: 175-185.
Greenbaum, D., Luscombe, N.M., Jansen, R., Qian, J., and Gerstein, M. (2001) Interrelating different types of genomic data, from proteome to secretome: 'oming in on function. Genome Res., 11: 1463-1468.
Huang, X., and Madan, A. (1999) CAP3: A DNA sequence assembly program. Genome Res., 9: 868-877.
Ilg, T., and Werr, M. (2012) Arginine kinase of the sheep blowfly Lucilia cuprina: Gene identification and characterization of the native and recombinant enzyme. Pesticide Biochemistry and Physiology, 102: 115-123.
Iwanaga, S., and Lee, B.L. (2005) Recent advances in the innate immunity of invertebrate animals. J. Biochem. Mol. Biol., 38: 128-150.
Jones, D.T., Taylor, W.R., and Thornton, J.M. (1992) The rapid generation of mutation data matrices from protein sequences. Comput. Appl. Biosci., 8: 275-282.
Lang, A.B., Wyss, C., and Eppenberger, H.M. (1980) Localization of arginine kinase in muscles fibres of Drosophila melanogaster. J. Muscle Res. Cell. Motil., 1: 147-161.
Liu, D.W. (2008) Opioid peptides and innate immune response in mollusc. Protein Pept. Lett., 15: 683-686.
Liu, E.T. (2005) Genomic technologies and the interrogation of the transcriptome. Mech. Ageing Dev., 126: 153-159.
McGuffin, L.J., Bryson, K., and Jones, D.T. (2000) The PSIPRED protein structure prediction server. Bioinformatics, 16: 404-405.
Mitta, G., Vandenbulcke, F., and Roch, P. (2000) Original involvement of antimicrobial peptides in mussel innate immunity. FEBS. Lett., 486: 185-190.
Newsholme, E.A., Beis, I., Leech, A.R., and Zammit, V.A. (1978) The role of creatine kinase and arginine kinase in muscle. Biochem. J., 172: 533-537.
Reddy, S.R., Houmeida, A., Benyamin, Y., and Roustan, C. (1992) Interaction in vitro of scallop muscle arginine kinase with filamentous actin. Eur. J. Biochem., 206: 251-257.
Rice, P., Longden, I., and Bleasby, A. (2000) EMBOSS: the European Molecular Biology Open Software Suite. Trends Genet., 16: 276-277.
Supajatura, V., Ushio, H., Nakao, A., Akira, S., Okumura, K., Ra, C., and Ogawa, H. (2002) Differential responses of mast cell Toll-like receptors 2 and 4 in allergy and innate immunity. J. Clin. Invest., 109: 1351-1359.
Suzuki, T., and Furukohri, T. (1994) Evolution of phosphagen kinase. Primary structure of glycocyamine kinase and arginine kinase from invertebrates. J. Mol. Biol., 237: 353-357.
Tamura, K., and Nei, M. (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol. Biol. Evol., 10: 512-526.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., and Kumar, S. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol., 28: 2731-2739.
van Deursen, J., Heerschap, A., Oerlemans, F., Ruitenbeek, W., Jap, P., ter Laak, H., and Wieringa, B. (1993) Skeletal muscles of mice deficient in muscle creatine kinase lack burst activity. Cell, 74: 621-631.
Wang, Y.E., Esbensen, P., and Bentley, D. (1998) Arginine kinase expression and localization in growth cone migration. J. Neurosci., 18: 987-998.
Wang, Z., Jian, J., Lu, Y., Wang, B., and Wu, Z. (2011) A tandem-repeat galectin involved in innate immune response of the pearl oyster Pinctada fucata. Mar. Genomics, 4: 229-236.
Yu, Z., He, X., Fu, D., and Zhang, Y. (2011) Two superoxide dismutase (SOD) with different subcellular localizations involved in innate immunity in Crassostrea hongkongensis. Fish Shellfish Immunol., 31: 533-539.
Zhang, H., Wang, L., Song, L., Song, X., Wang, B., Mu, C., and Zhang, Y. (2009) A fibrinogen-related protein from bay scallop Argopecten irradians involved in innate immunity as pattern recognition receptor. Fish Shellfish Immunol., 26: 56-64.
