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Tail-to-Head Tandem Duplication and Simple Repetitive Sequences of the Cytoplasmic Actin Genes in Greenling Hexagrammos otakii (Teleostei; Scorpaeniformes)

  • Lee, Sang-Yoon (Department of Marine Bio-Materials and Aquaculture, Pukyong National University) ;
  • Kim, Dong-Soo (Department of Marine Bio-Materials and Aquaculture, Pukyong National University) ;
  • Nam, Yoon-Kwon (Department of Marine Bio-Materials and Aquaculture, Pukyong National University)
  • Received : 2011.09.06
  • Accepted : 2011.11.29
  • Published : 2011.12.31

Abstract

We characterized a cytoplasmic actin gene locus in greenling Hexagrammos otakii (Scorpaeniformes). Genomic clones isolated from the greenling DNA library contained two homologous cytoplasmic actin gene copies (HObact2.1 and HObact2.2) in a tail-to-head orientation. Their gene structure is characterized by six translated exons and one non-translated exon. Exon-intron organization and the nucleotide sequences of the two actin gene isoforms are very similar. However, only the HObact2.1 isoform contains microsatellite-like, dinucleotide repeats in the 5'-flanking region (named HOms2002) and intron 1 following the non-translated exon 1 (named HOms769). One microsatellite locus (HOms769) was highly polymorphic while the other (HOms2002) was not. Based on bioinformatic analysis, different transcription factor binding motifs are related to stress and immune responses in the two actin isoforms. Semiquantitative and real-time reverse transcription-PCR assays showed that both isoform transcripts were detectable ubiquitously in all the tissues examined. However, the basal expression levels of each isoform varied across tissues. Overall, the two isoforms showed a similar, but not identical, expression pattern. Our data suggest that the cytoplasmic actin genes may be the result of a recent duplication event in the greenling genome, which has not experienced significant subfunctionalization in their housekeeping roles.

Keywords

References

  1. Anderson KV. 2000. Toll signaling pathways in the innate immune response. Curr Opin Immunol 12, 13-19. https://doi.org/10.1016/S0952-7915(99)00045-X
  2. Chaponnier C and Gabbiani G. 2004. Pathological situations characterized by altered actin isoform expression. J Pathol 204, 386-395. https://doi.org/10.1002/path.1635
  3. Chen SL, Miao GD, Shao CW, Tian YS and Liao X. 2009. Isolation and characterization of polymorphic microsatellite loci from fat greenling (Hexagrammos otakii). Conserv Genet 10, 1429-1431. https://doi.org/10.1007/s10592-008-9753-4
  4. Cho YS, Lee SY, Kim KH, Kim SK, Kim DS and Nam YK 2009. Gene structure and differential modulation of multiple rockbream (Oplegnathus fasciatus) hepcidin isoforms resulting from different biological stimulations. Dev Comp Immunol 33, 46-58. https://doi.org/10.1016/j.dci.2008.07.009
  5. Cho YS, Lee SY, Kim YK, Kim DS and Nam YK. 2011. Functional ability of cytoskeletal $\beta$-actin regulator to drive constitutive and ubiquitous expression of a fluorescent reporter throughout the life cycle of transgenic marine medaka Oryzias dancena. Transgenic Res 20, 1333-1355. https://doi.org/10.1007/s11248-011-9501-2
  6. Filby AL and Tyler CR. 2007. Appropriate 'housekeeping' genes for use in expression profiling the effects of environmental estrogens in fish. BMC Mol Biol 8, 10. https://doi.org/10.1186/1471-2199-8-10
  7. Franklin-Tong VE and Gourlay CW. 2008. A role for actin in regulating apoptosis/programmed cell death: evidence spanning yeast, plants and animals. Biochem J 413, 389-404. https://doi.org/10.1042/BJ20080320
  8. Jaillon O, Aury JM, Brunet F, Petit JL, Stange-Thomann N, Mauceli E, Bouneau L, Fischer C, Ozouf-Costaz C, Bernot A, Nicaud S, Jaffe D, Fisher S, Lutfalla G, Dossat C, Segurens B, Dasilva C, Salanoubat M, Levy M, Boudet N, Castellano S, Anthouard V, Jubin C, Castelli V, Katinka M, Vacherie B, Biemont C, Skalli Z, Cattolico L, Poulain J, De Berardinis V, Cruaud C, Duprat S, Brottier P, Coutanceau JP, Gouzy J, Parra G, Lardier G, Chapple C, McKernan KJ, McEwan P, Bosak S, Kellis M, Volff JN, Guigo R, Zody MC, Mesirov J, Lindblad-Toh K, Birren B, Nusbaum C, Kahn D, Robinson-Rechavi M, Laudet V, Schachter V, Quetier F, Saurin W, Scarpelli C, Wincker P, Lander ES, Weissenbach J, Roest Crollius H. 2004. Genome duplication in the teleost fish Tetraodon nigroviridis reveals the early vertebrate proto-karyotype. Nature 431, 946-957. https://doi.org/10.1038/nature03025
  9. Ji Y, Ferracci G, Warley A, Ward M, Leung KY, Samsuddin S, Leveque C, Queen L, Reebye V, Pal P, Gkaliagkousi E, Seager M and Ferro A. 2007. $\beta$-Actin regulates platelet nitric oxide synthase 3 activity through interaction with heat shock protein 90. Proc Natl Acad Sci U S A 104, 8839-8844. https://doi.org/10.1073/pnas.0611416104
  10. Kang HW, Chung EY and Kim JH. 2004. Sexual maturation and spawning characteristics in greenling Hexagrammos otakii of the west coast in Korea. J Aquac 17, 30-38.
  11. Kim KY, Lee SY, Cho YS, Bang IC, Kim DS and Nam YK. 2008. Characterization and phylogeny of two $\beta$-cytoskeletal actins from Hemibarbus mylodon (Cyprinidae, Cypriniformes), a threatened fish species in Korea. DNA Seq 19, 87-97. https://doi.org/10.1080/10425170701445691
  12. Kim Y, Kim PK, Kim HJ, Jo JY, Han W and Park J. 2009. Effects of water temperature and ambient ammonia concentration on oxygen consumption and ammonia excretion of greenling Hexagrammos otakii Jordan et Starks. Korean J Fish Aquat Sci 42, 373-379. https://doi.org/10.5657/kfas.2009.42.4.373
  13. Kubista M, Andrade JM, Bengtsson M, Forootan A, Jonak J, Lind K, Sindelka R, Sjoback R, Sjogreen B, Strombom L, Stahlberg A and Zoric N. 2006. The real-time polymerase chain reaction. Mol Aspects Med 27, 95-125. https://doi.org/10.1016/j.mam.2005.12.007
  14. Kustermans G, Piette J and Legrand-Poels S. 2008. Actin-targeting nating ural compounds as tools to study the role of actin cytoskeleton in signal transduction. Biochem Pharmacol 76, 1310-1322. https://doi.org/10.1016/j.bcp.2008.05.028
  15. Lee SY, Kim KH and Nam YK. 2009. Molecular characterization of rockbream (Oplegnathus fasciatus) cytoskeletal $\beta$-actin gene and its 5'-upstream regulatory region. Fish Aquat Sci 12, 90-97. https://doi.org/10.5657/fas.2009.12.2.090
  16. Liu ZJ and Cordes JF. 2004. DNA marker technologies and their applications in aquaculture genetics. Aquaculture 238, 1-37. https://doi.org/10.1016/j.aquaculture.2004.05.027
  17. Liu ZJ, Moav B, Faras AJ, Guise KS, Kapuscinski AR and Hackett P. 1991. Importance of the CArG box in regulation of $\beta$-actin-encoding genes. Gene 108, 211-217. https://doi.org/10.1016/0378-1119(91)90436-F
  18. Miwa T, Manabe Y, Kurokawa K, Kamada S, Kanda N, Bruns G, Ueyama H and Kakunaga T. 1991. Structure, chromosome location, and expression of the human smooth muscle (enteric type) $\gamma$-actin gene: evolution of six human actin genes. Mol Cell Biol 11, 3296-3306.
  19. Nam YK and Kim DS. 2002. Molecular cloning and alternative splicing of growth hormone transcripts in greenling, Hexagrammos otakii. J Korean Fish Soc 35, 676-681.
  20. Nam YK, Maclean N, Hwang G and Kim DS. 2008. Autotransgenic and allotransgenic manipulation of growth traits in fish for aquaculture: a review. J Fish Biol 72, 1-26. https://doi.org/10.1111/j.1095-8649.2007.01738.x
  21. Noh JK, Cho KN, Han EH, Kim A, Lee JS, Kim DS and Kim CG. 2003. Genomic cloning of mud loach Misgurnus mizolepis (Cyprini formes, Cobitidae) $\beta$-actin gene and usefulness of its promoter region for fish transgenesis. Mar Biotechnol 5, 244-252. https://doi.org/10.1007/s10126-002-0066-1
  22. Percipalle P and Visa N. 2006. Molecular functions of nuclear actin in transcription. J Cell Biol 172, 967-971. https://doi.org/10.1083/jcb.200512083
  23. Reece KS, McElroy D and Wu R. 1992. Function and evolution of actins. Evol Biol 26, 1-34.
  24. Reisler E and Egelman EH. 2007. Actin structure and function: what we still do not understand. J Biol Chem 282, 36133-36137. https://doi.org/10.1074/jbc.R700030200
  25. Schmittgen TD and Livak KJ. 2008. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3, 1101-1108. https://doi.org/10.1038/nprot.2008.73
  26. Small BC, Murdock CA, Bilodeau-Bourgeois AL, Peterson BC and Waldbieser GC. 2008. Stability of reference genes for real-time PCR analyses in channel catfish (Ictalurus punctatus) tissues under varying physiological conditions. Comp Biochem Physiol B Biochem Mol Biol 151, 296-304. https://doi.org/10.1016/j.cbpb.2008.07.010
  27. Truksa J, Lee P and Beutler E. 2007. The role of STAT, AP-1, E-box and TIEG motifs in the regulation of hepcidin by IL-6 and BMP-9: lessons from human HAMP and murine Hamp1 and Hamp2 gene promoters. Blood Cells Mol Dis 39, 255-262. https://doi.org/10.1016/j.bcmd.2007.06.014
  28. Venkatesh B, Tay BH, Elgar G and Brenner S. 1996. Isolation, characterization and evolution of nine pufferfish (Fugu rubripes) actin genes. J Mol Biol 259, 655-665. https://doi.org/10.1006/jmbi.1996.0347