ISSN : 1225-3480
Metallothioneins (MTs) play a key role in metallic homeostasis and detoxification in most living organisms. In an attempt to study the biological functions and significance of MT in a snail, we cloned and partially characterized the MT gene from the left-handed snail, Physa acutawhich has been regarded as a potential biomonitering species for fresh water. The complete cDNA sequence of PaMT cDNA was identified from the expressed sequence tag (EST) sequencing project of Physa acuta. The coding region of 180 bp gives 60 amino acid residues including the initiation methionine and termination codon. Clustering and phylogenic analysis of PaMT with other MT amino acid sequences show that it has some identities to Helix pomatia (60%), Arianta arbustorum (58%), Perna viridis (49%), Mytilus edulis (49%), Bathymodiolus azoricus (49%), Bathymodiolus azoricus (48%) and Bathymodiolus sp. FD-2002 (48%). Time dependent induction for PaMT from P. acuta exposed with cadmium (50 ppb) indicated that PaMT was induced at 4-8 hr after exposure. It remains to further develop PaMTas a potential biomarker for water contamination in fresh water.
Altschul, S.F., Gish, W., Miller, W., Myers, E.W., and Lipman, D.J. (1990) Basic local alignment search tool. Journal of molecular biology 215: 403-410.
Azevedo, J.S., Serafim, A., Company, R., Braga, E.S., Favaro, D.I., and Bebianno, M.J. (2009) Biomarkers of exposure to metal contamination and lipid peroxidation in the benthic fish Cathorops spixii from two estuaries in South America, Brazil. Ecotoxicology 18: 1001-1010.
Bacchetta, R., Mantecca, P., and Vailati, G. (2002) Oocyte Degeneration and Altered Ovipository Activity Induced by Paraquat in the Freshwater Snail Physa Fontinalis (Gastropoda: Pulmonata). Journal of Molluscan Study 68: 181-186.
Baek, M.K., Lee, J.-S., Kang, S.W., Lee, J.B., Kang, H.J., Jo, Y.-H., Noh, M.-Y., Han, Y.S., Choi, S.-H., Chae, S.-H., Park, H.-S., Lee, J.-S., and Lee, Y.S. (2009) Phylogenetic analysis based on metallothionein gene sequence of an indigenous species Pisidium (Neopisidium) coreanum in Korea. Korean Journal of Malacology 25: 153-160.
Bebianno, M.J., and Serafim, M.A. (2003) Variation of metal and metallothionein concentrations in a natural population of Ruditapes decussatus. Archives of environmental contamination and toxicology 44: 53-66.
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.
Butler, R.A., and Roesijadi, G. (2001) Quantitative reverse transcription polymerase chain reaction of a molluscan metallothionein mRNA. Aquatic toxicology 54: 59-67.
Canivet, V., Chambon, P., and Gibert, J. (2001) Toxicity and bioaccumulation of arsenic and chromium in epigean and hypogean freshwater macroinvertebrates. Archives of environmental contamination and toxicology 40: 345-354.
Choi, H.J., Ahn, I.-Y., Ryu, S.-K., Lee, Y.-S., Lee, I.-S., and Jeong, K.-H. (2001) Preliminary Evidence for a Metallothionein-like Cd-binding Protein in the Kidney of the Antarctic Clam Laternula elliptica. Ocean and Polar Research 23: 337-345.
Cuppen, J.G., Crum, S.J., Van den Heuvel, H.H., Smidt, R.A., and Van den Brink, P.J. (2002) Effects of a mixture of two insecticides in freshwater microcosms: I. Fate of chlorpyrifos and lindane and responses of macroinvertebrates. Ecotoxicology 11: 165-180.
Dopazo, J. (1994) Estimating errors and confidence intervals for branch lengths in phylogenetic trees by a bootstrap approach. Journal of molecular evolution 38: 300-304.
Ewing, B., and Green, P. (1998) Base-calling of automated sequencer traces using phred. II. Error probabilities. Genome Research 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 Research 8: 175-185.
Felsenstein, J. (1985) Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39: 783-791.
Huang, X., and Madan, A. (1999) CAP3: A DNA sequence assembly program. Genome Research 9: 868-877.
Johnson, M., Zaretskaya, I., Raytselis, Y., Merezhuk, Y., McGinnis, S., and Madden, T.L. (2008) NCBI BLAST: a better web interface. Nucleic Acids Research 36: W5-9.
Kagi, J.H., and Valee, B.L. (1960) Metallothionein: a cadmium- and zinc-containing protein from equine renal cortex. The Journal of biological chemistry 235: 3460-3465.
Kagi, J.H., and Vallee, B.L. (1961) Metallothionein: a cadmium and zinc-containign protein from equine renal cortex. II. Physico-chemical properties. The Journal of biological chemistry 236: 2435-2442.
Kojima, Y., Berger, C., Vallee, B.L., and Kagi, J.H. (1976) Amino-acid sequence of equine renal metallothionein-1B. Proceedings of the National Academy of Sciences of the United States of America 73: 3413-3417.
Lee, J.-S., Min, B.-J., Kang, S.W., Lee, J.B., Baek, M.K., Hwang, S.Y., Kim, S.H., Kho,W.-G., Choi, S.-H., Chae, S.-H., Park, H.-S., Han, Y.S., Lee, J.-S., Jeong, K.-H., and Lee, Y.S. (2008) Molecular Phylogenetic Study of Nesiohelix samarangae Based on Metallothionein Gene. Korean Journal of Malacology 24: 73-80.
Lee, Y.-S., Jo, Y.-H., Han, Y.-S., Kho, W.-G., Ahn, I.-Y., and Jeong, K.-H. (2006) Bioaccumulation of Heavy Metals in Intestine of Nacella concinna. Korean Journal of Malacology 22: 83-91.
Lee, Y.-S., Jo, Y.-H., Kim, D.-S., Kim, D.-W., Kim, M.-Y., Choi, S.-H., Yon, J.-O., Byun, I.-S., Kang, B.-R., Jeong, K.-H., and Park, H.-S. (2004) Construction of BLAST Server for Mollusks. Korean Journal of Malacology 20: 165-169.
Lee, Y.S., Ahn, I.-Y., Kim, W.J., and Jeong, K.H. (2002) An immunohistochemical and ultrastructural study of the heavy metals accumulated in the kidney of an antarctic clam, Laternula elliptica. Korean Journal of Malacology 18: 15-21.
Lee, Y.S., Jo, Y.H., Byun, I.S., Kang, S.W., Cho, E.M., Han, Y.S., Choi, S.-H., Park, H.-S., Kho, W.-G., Ahn, I.-Y., and Jeong, K.-H. (2006) Bioaccumulation of Heavy Metals in Ruditapes philippinarum. Korean Journal of Malacology 22: 157-165.
Leung, K.M.Y., Morley, N.J., Grist, E.P.M., Morritt, D., and Crane, M. (2004) Chronic toxicity of tributyltin on development and reproduction of the hermaphroditic snail Physa fontinalis: Influence of population density. Marine Environmental Research 58: 157-162.
Maltez, H.F., Villanueva Tagle, M., Fernandez de la Campa Mdel, R., and Sanz-Medel, A. (2009) Metal-metallothioneins like proteins investigation by heteroatom-tagged proteomics in two different snails as possible sentinel organisms of metal contamination in freshwater ecosystems. Analytica chimica acta 650: 234-240.
Mourgaud, Y., Martinez, E., Geffard, A., Andral, B., Stanisiere, J.Y., and Amiard, J.C. (2002) Metallothionein concentration in the mussel Mytilus galloprovincialis as a biomarker of response to metal contamination: validation in the field. Biomarkers 7: 479-490.
Nebeker, A.V., and Schuytema, G.S. (1998) Chronic effects of the herbicide diuron on freshwater cladocerans, amphipods, midges, minnows, worms, and snails. Archives of environmental contamination and toxicology 35: 441-446.
OConnor, T.P., Cantillo, A.Y., and Lauenstein, G.G. (1994) Monitoring of Temporal trends in chemical contamination by the NOAA National Status and Trends Mussel Watch Project. In: Biomonitoring of Costal Waters and Estuaries. (ed. by Kramer, K.J.M.)
Saitou, N., and Nei, M. (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular biology and evolution 4: 406-425.
Simes, D.C., Bebianno, M.J., and Moura, J.J. (2003) Isolation and characterisation of metallothionein from the clam Ruditapes decussatus. Aquatic toxicology 63: 307-318.
Stobaeus, J.K., Heath, G.E., Parkhurst, R.M., Jones, W.O., and Webster, J.E. (1990) A laboratory study of the toxicity of the butanol extract of endod (Phytolacca dodecandra) on two species of freshwater fish and two species of aquatic snails. Veterinary & Human Toxicology 32: 212-216.
Takimoto, Y., Ohshima, M., and Miyamoto, J. (1987) Comparative metabolism of fenitrothion in aquatic organisms. II. Metabolism in the freshwater snails, Cipangopaludina japonica and Physa acuta. Ecotoxicology and environmental safety 13: 118-125.
Tamura, K., Dudley, J., Nei, M., and Kumar, S. (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular biology and evolution 24: 1596-1599.
Tanguy, A., and Moraga, D. (2001) Cloning and characterization of a gene coding for a novel metallothionein in the Pacific oyster Crassostrea gigas (CgMT2): a case of adaptive response to metal-induced stress? Gene 273: 123-130.
Tanguy, A., Mura, C., and Moraga, D. (2001) Cloning of a metallothionein gene and characterization of two other cDNA sequences in the Pacific oyster Crassostrea gigas (CgMT1). Aquatic toxicology 55: 35-47.
Zdobnov, E.M., and Apweiler, R. (2001) InterProScan--an integration platform for the signature-recognition methods in InterPro. Bioinformatics 17: 847-848.
Zuckerkandl, E., and Pauling, L. (1965) Evolutionary divergence and convergence in proteins pp. Academic Press. New York
