Identification and characterization of a diamine exporter in colon epithelial cells

Takeshi Uemura, Hagit F. Yerushalmi, George Tsaprailis, David E. Stringer, Kirk E. Pastorian, Leo Hawel, Craig V. Byus, Eugene W. Gerner

Research output: Contribution to journalArticle

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Abstract

SLC3A2, a member of the solute carrier family, was identified by proteomics methods as a component of a transporter capable of exporting the diamine putrescine in the Chinese hamster ovary (CHO) cells selected for resistance to growth inhibition by high exogenous concentrations of putrescine. Putrescine transport was increased in inverted plasma membrane vesicles prepared from cells resistant to growth inhibition by putrescine compared with transport in inverted vesicles prepared from non-selected cells. Knockdown of SLC3A2 in human cells, using short hairpin RNA, caused an increase in putrescine uptake and a decrease in arginine uptake activity. SLC3A2 knockdown cells accumulated higher polyamine levels and grew faster than control cells. The growth of SLC3A2 knockdown cells was inhibited by high concentrations of putrescine. Knockdown of SLC3A2 reduced export of polyamines from cells. Expression of SLC3A2 was suppressed in human HCT116 colon cancer cells, which have an activated K-RAS, compared with their isogenic clone, Hkh2 cells, which lack an activated K-RAS allele. Spermidine/spermine N1-acetyltransferase (SAT1) was co-immunoprecipitated by an anti-SLC3A2 antibody as was SLC3A2 with an anti-SAT1 antibody. SLC3A2 and SAT1 colocalized on the plasma membrane. These data provide the first molecular characterization of a polyamine exporter in animal cells and indicate that the diamine putrescine is exported by an arginine transporter containing SLC3A2, whose expression is negatively regulated by K-RAS. The interaction between SLC3A2 and SAT1 suggests that these proteins may facilitate excretion of acetylated polyamines.

Original languageEnglish (US)
Pages (from-to)26428-26435
Number of pages8
JournalJournal of Biological Chemistry
Volume283
Issue number39
DOIs
StatePublished - Sep 26 2008

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Diamines
Putrescine
Colon
Epithelial Cells
Polyamines
Cells
Cell membranes
Arginine
Clone cells
Acetyltransferases
Anti-Idiotypic Antibodies
Spermidine
Spermine
Antibodies
Growth
Cell Membrane
Small Interfering RNA
Animals
Cricetulus
Proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Uemura, T., Yerushalmi, H. F., Tsaprailis, G., Stringer, D. E., Pastorian, K. E., Hawel, L., ... Gerner, E. W. (2008). Identification and characterization of a diamine exporter in colon epithelial cells. Journal of Biological Chemistry, 283(39), 26428-26435. https://doi.org/10.1074/jbc.M804714200

Identification and characterization of a diamine exporter in colon epithelial cells. / Uemura, Takeshi; Yerushalmi, Hagit F.; Tsaprailis, George; Stringer, David E.; Pastorian, Kirk E.; Hawel, Leo; Byus, Craig V.; Gerner, Eugene W.

In: Journal of Biological Chemistry, Vol. 283, No. 39, 26.09.2008, p. 26428-26435.

Research output: Contribution to journalArticle

Uemura, T, Yerushalmi, HF, Tsaprailis, G, Stringer, DE, Pastorian, KE, Hawel, L, Byus, CV & Gerner, EW 2008, 'Identification and characterization of a diamine exporter in colon epithelial cells', Journal of Biological Chemistry, vol. 283, no. 39, pp. 26428-26435. https://doi.org/10.1074/jbc.M804714200
Uemura, Takeshi ; Yerushalmi, Hagit F. ; Tsaprailis, George ; Stringer, David E. ; Pastorian, Kirk E. ; Hawel, Leo ; Byus, Craig V. ; Gerner, Eugene W. / Identification and characterization of a diamine exporter in colon epithelial cells. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 39. pp. 26428-26435.
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