title: Characterization of the selenite uptake mechanism in the coccolithophore Emiliania huxleyi (Haptophyta).
abstract: The marine coccolithophore Emiliania huxleyi (Haptophyta) requires selenium as an essential element for growth, and the active species absorbed is selenite, not selenate. This study characterized the selenite uptake mechanism using ⁷⁵Se as a tracer. Kinetic analysis of selenite uptake showed the involvement of both active and passive transport processes. The active transport was suppressed by 0.5 mM vanadate, a membrane-permeable inhibitor of H⁺-ATPase, at pH 8.3. When the pH was lowered from 8.3 to 5.3, the selenite uptake activity greatly increased, even in the presence of vanadate, suggesting that the H⁺ concentration gradient may be a motive force for selenite transport. [⁷⁵Se]Selenite uptake at selenite-limiting concentrations was hardly affected by selenate, sulfate and sulfite, even at 100 μM. In contrast, 3 μM orthophosphate increased the K(m) 5-fold. These data showed that HSeO₃⁻, a dominant selenite species at acidic pH, is the active species for transport through the plasma membrane and transport is driven by ΔpH energized by H⁺-ATPase. Kinetic analysis showed that the selenite uptake activity was competitively inhibited by orthophosphate. Furthermore, the active selenite transport mechanism was shown to be induced de novo under Se-deficient conditions and induction was suppressed by the addition of either sufficient selenite or cycloheximide, an inhibitor of de novo protein synthesis. These results indicate that E. huxleyi cells developed an active selenite uptake mechanism to overcome the disadvantages of Se limitation in ecosystems, maintaining selenium metabolism and selenoproteins for high viability.
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba 305-8572, Japan.
authors.forename: Hiroya
authors.initials: H
authors.lastname: Araie
authors.forename: Kou
authors.initials: K
authors.lastname: Sakamoto
authors.forename: Iwane
authors.initials: I
authors.lastname: Suzuki
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Shiraiwa
mesh_terms: D001693:Biological Transport, Active; D058087:Haptophyta / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism; D064586:Selenic Acid; D018036:Selenium Compounds / Q000378:metabolism; D018038:Sodium Selenite / Q000493:pharmacokinetics*; D013447:Sulfites / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology
chemical_list: D018036:Selenium Compounds; D013447:Sulfites; D014638:Vanadates; D006180:Proton-Translocating ATPases; D018038:Sodium Selenite; D064586:Selenic Acid