The expression of the open reading frame of Arabidopsis CAX1, but not its cDNA, confers metal tolerance in yeast.
The biochemical properties and regulation of several plant CAX (CAtion eXchanger)-type vacuolar Ca2+/H+ exchangers have been extensively analysed in yeast expression assays. In the present study, we compare and contrast the phenotypes of yeast cells expressing the CAX1 cDNA and open reading frame (ORF). We report that the CAX1 ORF, but not the cDNA containing the 3′-untranslated region (UTR), was able to confer Ca2+ tolerance when expressed in a Ca2+-sensitive yeast mutant. Additionally, only yeasts expressing the N-terminal truncated CAX1 ORF were able to grow on high Mn2+ media, suggesting that removal of the 3′-UTR altered activity. However, removal of the 3′-UTR from another CAX did not alter the yeast phenotypes. Expression studies demonstrated that expressing the CAX1 ORF in yeast elevates CAX1 RNA and protein levels. Our results suggest that the 3′-UTR modulates expression of CAX1 in yeast.
United States Department of Agriculture ⁄Agricultural Research Service Children’s Nutrition Research Center, Baylor College of Medicine, Houston, TX, USA. toshiro.shigaki@nari.org.pg
T
T
Shigaki
H
H
Mei
J
J
Marshall
X
X
Li
M
M
Manohar
K D
KD
Hirschi
D020413:3' Untranslated Regions; D017920:Antiporters / Q000235:genetics*; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics*; D002118:Calcium / Q000378:metabolism; D027682:Cation Transport Proteins / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D016366:Open Reading Frames; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
D020413:3' Untranslated Regions; D017920:Antiporters; D029681:Arabidopsis Proteins; D027682:Cation Transport Proteins; D018076:DNA, Complementary; C027614:calcium-hydrogen antiporters; D002118:Calcium