Expressed sequence tag analysis of vanadocytes in a vanadium-rich ascidian, Ascidia sydneiensis samea.
Some species in the family Ascidiidae accumulate vanadium at concentrations in excess of 350 mM, which corresponds to about 10(7) times higher than that in seawater. In these species signet ring cells, with a single huge vacuole in which vanadium ion is contained, function as vanadium-accumulating cells, vanadocytes. To investigate the mechanism underlying this phenomenon, we performed an expressed sequence tag (EST) analysis of a complementary DNA library from vanadocytes of a vanadium-rich ascidian, Ascidia sydneiensis samea. We determined the nucleotide sequences of 1000 ESTs and performed a BLAST analysis against the SwissProt database. We found 93 clones of metal-related gene homologues, including the ferritin heavy subunit, hemocyanin, and metallothionein. Two ESTs, in particular, exhibited significant similarity to vanabins that have been extracted from A. sydneiensis samea blood cells as low molecular weight vanadium-binding proteins. We have named the genes encoding these ESTs vanabin3 and vanabin4. Immobilized metal ion affinity chromatography revealed that these novel vanabin homologues bind vanadium(IV) ions.
Yamaguchi
Nobuo
N
Marine Biological Laboratory, Graduate School of Science, Hiroshima University, Mukaishima-cho 2445, Hiroshima 722-0073, Japan.
Kamino
Kei
K
Ueki
Tatsuya
T
Michibata
Hitoshi
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002846:Chromatography, Affinity; D019295:Computational Biology; D017931:DNA Primers; D020224:Expressed Sequence Tags; D005293:Ferritins / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D006433:Hemocyanins / Q000235:genetics / Q000378:metabolism; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D014561:Urochordata / Q000235:genetics*; D014617:Vacuoles / Q000737:chemistry / Q000235:genetics*; D014639:Vanadium / Q000378:metabolism*
D017931:DNA Primers; D014639:Vanadium; D005293:Ferritins; D006433:Hemocyanins; D008668:Metallothionein