Marine Biotechnology

, Volume 7, Issue 6, pp 677–686 | Cite as

Structural Analysis of cDNAs Coding for 4SNc-Tudor Domain Protein from Fish and Their Expression in Yellowtail Organs

  • Shunnosuke AbeEmail author
  • Pi-Lin Wang
  • Fuminori Takahashi
  • Eiji Sasaki


We cloned complementary DNAs for 4SNc-Tudor protein (SN4TDR) from yellowtail (Seriola quinqueradiata), torafugu (Takifugu rubripes), and zebrafish (Danio rerio). This protein contains 4 staphylococcal nuclease domains at the N terminus followed by a Tudor domain. We also identified the 4SNc-Tudor proteins highly homologous to that in yellowtail from the Takifugu genomic database. According to the smart database, these fish proteins had an overlapping Tudor domain (smart00333) with a complete 5 SNc domain (smart00318). In addition, 2 possible translation start sites were observed at the 5′ sequences in all 3 fish species. Northern blot analysis of different yellowtail organs showed that the full SN4TDR messenger RNA was approximately 4000 nucleotides long and that its expression was highest in liver and gallbladder, being about 2 to 5 times higher than in kidney, brain, ovary, and gills, and exceedingly low in spleen, heart, and muscle. A minor 2000-nucleotide transcript observed in kidney, spleen, and gallbladder, was attributable to an alternatively spliced variant of this gene. Total proteins extracted from yellowtail liver were fractionated by heparin affinity column chromatography and separated by sodium dodecylsulfate polyacrylamide gel electrophoresis. Analyses by SDS-PAGE and liquid chromatography with tandem mass spectroscopy identified the polypeptide encoded by SN4TDR as a single molecule of 102 kDa.


SN4TDR SND1 LRRC4 proteomics torafugu zebrafish 


  1. Abe S, Sakai M, Yagi K, Hagino T, Ochi K, Shibata K, Davies E (2003) A Tudor protein with multiple SNc domains from pea seedlings: cellular localization, partial characterization, sequence analysis, and phylogenetic relationships. J Exp Bot 54: 971–983CrossRefGoogle Scholar
  2. Abe S, Chamnan C, Miyamoto K, Minamino Y, Nouda M (2004a) Isolation and identification of 3-methylcrotonyl coenzyme A carboxylase cDNAs and pyruvate carboxylase, and their expression in red seabream (Pagrus major) organs. Mar Biotechnol 6, 527–540CrossRefGoogle Scholar
  3. Abe S, Chiba S, Mishra N, Minamino Y, Nakasuji H, Doi M, Gray TA (2004) Origin and evolution of the genomic region encoding RAF1, MKRN2, PPARG, and SYN2 in human chromosome 3p25. Mar Biotechnol 6: S404–S412CrossRefGoogle Scholar
  4. Abe S, Davies E (1991) Isolation of F-actin from pea stems: Evidence from fluorescence microscopy. Protoplasma 163: 51–61CrossRefGoogle Scholar
  5. Broadhurst MK, Wheeler TT (2001) The p100 coactivator is present in the nuclei of mammary epithelial cells and its abundance is increased in response to prolactin in culture and in mammary tissue during lactation. Journal of Endocrinology 171: 329–337CrossRefGoogle Scholar
  6. Caudy AA, Ketting RF, Hammond SM, Denli AM, Bathoorn AM, Tops BB, Silva JM, Myers MM, Hannon GJ, Plasterk RH (2003) A micrococcal nuclease homologue in RNAi effector complexes. Nature 425(6956): 411–414CrossRefGoogle Scholar
  7. Chamnan C, Abe S, Doi M, Chiba S, Gray TA (2003) The genomic organization of MKRN1, and expression profile of MKRN1, MKRN2, and RAF1 in yellowtail fish (Seriola quinqueradiata). Histol Histochem 42: 57–75Google Scholar
  8. Davies E, Stankovic B, Azama K, Shibata K, Abe S (2001) Novel components of the plant cytoskeleton: A beginning to plant ‘cytomics’. Plant Science 160: 185–196CrossRefGoogle Scholar
  9. Ito Y, Abe S, Davies E (1994) Co-localization of cytoskeleton proteins and polysomes with a membrane fraction from peas. Journal of Experimental Botany 45: 253–359Google Scholar
  10. Gray TA, Azama K, Whitmore K, Min A, Abe S, Nicholls RD (2001) Phylogenetic conservation of the makorin-2 gene, encoding multiple zinc-finger protein,antisense to the RAF1 proto-oncogene. Genomics 77: 119–126CrossRefGoogle Scholar
  11. Keefe LJ, Sondek J, Shortle D, Lattman EE (1993) The alpha aneurism: a structural motif revealed in an insertion mutant of staphylococcal nuclease. Proceedings of the National Academy of Sciences of the United States of America 90: 3275–3279Google Scholar
  12. Keefe LJ, Quirk S, Gittis A, Sondek J, Lattman EE (1994) Accommodation of insertion mutations on the surface and in the interior of staphylococcal nuclease. Protein Science 3: 391–401Google Scholar
  13. Kozak M (1999) Initiation of translation in prokaryotes and eukaryotes. Gene 2: 187–208Google Scholar
  14. Liu Z, Kim S, Karsi A (2001) Channel catfish follicle-stimulating hormone and luteinizing hormone: Complementary DNA cloning and expression during ovulation. Mar Biotechnol 3: 590–599CrossRefGoogle Scholar
  15. Peyush P, Moriyama S, Takahashi A, Kawauchi H (2000) Molecular cloning of growth hormone complementary DNA in barfin flounder (Verasper moseri). Mar Biotechnol 2: 21–26Google Scholar
  16. Pohar N, Godenschwege TA, Buchner E (1999) Invertebrate tissue inhibitor of metalloproteinase: Structure and nested gene organization within the synapsin locus is conserved from Drosophila to human. Genomics 57(2): 293–296CrossRefGoogle Scholar
  17. Ponting CP (1997) Tudor domains in proteins that interact with RNA. Trends Biochem Sci 2: 51–52Google Scholar
  18. Porta A, Colonna-Romano S, Callebaut I, Franco A, Marzullo L, Kobayashi GS, Maresca B (1999) An homologue of the human 100-kDa protein (p100) is differentially expressed by Histoplasma capsulatum during infection of murine macrophages. Biochem Biophys Res Commun 254: 605–613CrossRefGoogle Scholar
  19. Sami-Subbu R, Choi S-B, Wu Y, Wang C, Okita TW (2001) Identification of a cytoskeleton-associated 120 kDa RNA-binding protein in developing rice seeds. Plant Mol Biol 46: 79–88CrossRefGoogle Scholar
  20. Shibata K, Morita Y, Abe S, Stankovic B, Davies E (1999) Apyrase from pea stems: Isolation, purification, characterization and identification of a NTPase from the cytoskeleton fraction of pea stem tissue. Plant Physiol Biochemistry 37: 881–888CrossRefGoogle Scholar
  21. Strausberg RL, Feingold EA, Marra MA (2002) Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences. J Proc Natl Acad Sci USA 99(26): 16899–16903Google Scholar
  22. Tijsterman M, May RC, Simmer F, Okihara KL, Plasterk RH (2004). Genes required for systemic RNA interference in Caenorhabditis elegans. Current Biology 14(2): 111–116CrossRefGoogle Scholar
  23. Tong X, Drapkin R, Yalamanchili R, Mosialos G, Kieff E (1995) The Epstein-Barr virus nuclear protein 2 acidic domain forms a complex with a novel cellular coactivator that can interact with TFIIE. Mol Cell Biol 15: 4735–4744Google Scholar
  24. Wang JR, Li XL, Fan SQ, Tan C, Xiang JJ, Tang K, Wang R, Li GY (2003) Expression of LRRC4 has the potential to decrease the growth rate and tumorigenesis of glioblastoma cell line U251. Ai Zheng. 22(9): 897–902Google Scholar
  25. Yang J, Aittomäki S, Pesu M, Carter K, Saarinen J, Kalkkinen N, Kieff E, Silvennoinen O (2002) Identification of p100 as a coactivator for STAT6 that bridges STAT6 with RNA polymerase II. EMBO J 21(18): 4950–4958CrossRefGoogle Scholar
  26. Zhao CT, Shi KH, Su Y, Liang LY, Yan Y, Postlethwait J Meng AM (2003) Two variants of zebrafish p100 are expressed during embryogenesis and regulated by Nodal signaling. FEBS Lett 543(1–3): 190–195Google Scholar

Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Shunnosuke Abe
    • 1
    Email author
  • Pi-Lin Wang
    • 1
  • Fuminori Takahashi
    • 1
  • Eiji Sasaki
    • 1
  1. 1.Laboratory of Molecular Cell Biology, Faculty of AgricultureEhime UniversityMatsuyamaJapan

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