A novel plant vacuolar Na+/H+ antiporter gene evolved by DNA shuffling confers improved salt tolerance in yeast.
Plant vacuolar Na(+)/H(+) antiporters play important roles in maintaining cellular ion homeostasis and mediating the transport of Na(+) out of the cytosol and into the vacuole. Vacuolar antiporters have been shown to play significant roles in salt tolerance; however the relatively low V(max) of the Na(+)/H(+) exchange of the Na(+)/H(+) antiporters identified could limit its application in the molecular breeding of salt tolerant crops. In this study, we applied DNA shuffling methodology to generate and recombine the mutations of Arabidopsis thaliana vacuolar Na(+)/H(+) antiporter gene AtNHX1. Screening using a large scale yeast complementation system identified AtNHXS1, a novel Na(+)/H(+) antiporter. Expression of AtNHXS1 in yeast showed that the antiporter localized to the vacuolar membrane and that its expression improved the tolerance of yeast to NaCl, KCl, LiCl, and hygromycin B. Measurements of the ion transport activity across the intact yeast vacuole demonstrated that the AtNHXS1 protein showed higher Na(+)/H(+) exchange activity and a slightly improved K(+)/H(+) exchange activity.
Xu
Kai
K
Department of Biochemistry and Molecular Biology, School of Life Science, East China Normal University, Shanghai 200062, China.
Zhang
Hui
H
Blumwald
Eduardo
E
Xia
Tao
T
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000166:cytology / Q000235:genetics*; D032621:DNA Shuffling; D004305:Dose-Response Relationship, Drug; D017343:Genes, Plant / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D021381:Protein Transport; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D055049:Salt Tolerance / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D012964:Sodium / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D017923:Sodium-Hydrogen Exchangers / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
D017923:Sodium-Hydrogen Exchangers; D012965:Sodium Chloride; D012964:Sodium
Mol Cell Biochem. 2003 Dec;254(1-2):117-24
14674689
J Biol Chem. 2003 Jun 20;278(25):22453-9
12695519
FEBS Lett. 2003 Jun 19;545(2-3):239-45
12804783
Biochem J. 2001 Jul 1;357(Pt 1):1-10
11415429
Mol Membr Biol. 2006 Jul-Aug;23(4):349-61
16923728
Nat Biotechnol. 2001 Aug;19(8):765-8
11479571
Nature. 1998 Jan 15;391(6664):288-91
9440693
J Mol Biol. 1988 Jun 5;201(3):601-19
3418712
Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10747-51
7938023
FEBS Lett. 2000 Apr 14;471(2-3):224-8
10767428
J Biochem. 2002 Jun;131(6):821-31
12038978
Nat Biotechnol. 2003 Jan;21(1):81-5
12469134
Plant J. 2003 Oct;36(2):229-39
14535887
Channels (Austin). 2008 Sep-Oct;2(5):329-36
19001864
Mol Biol Cell. 2000 Dec;11(12):4277-94
11102523
FEBS Lett. 2007 May 25;581(12):2247-54
17459382
Mol Microbiol. 2001 May;40(3):656-68
11359571
Nucleic Acids Res. 1997 Mar 15;25(6):1307-8
9092645
Nature. 1994 Aug 4;370(6488):389-91
8047147
J Bacteriol. 1984 Sep;159(3):940-5
6384187
Proc Natl Acad Sci U S A. 2005 Nov 1;102(44):16107-12
16249341
J Biol Chem. 2007 Dec 28;282(52):37854-63
17981808
J Biol Chem. 1998 Jun 12;273(24):14838-44
9614085
Biochem J. 2000 Oct 1;351(Pt 1):241-9
10998367
Nucleic Acids Res. 1992 Mar 25;20(6):1425
1561104
Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12832-6
11606781
Plant J. 2004 Dec;40(5):752-71
15546358
Trends Plant Sci. 2005 Dec;10(12):615-20
16280254
Science. 1999 Aug 20;285(5431):1256-8
10455050
J Biol Chem. 2002 Jan 25;277(4):2413-8
11707435
Biochim Biophys Acta. 2000 May 1;1465(1-2):140-51
10748251
J Biol Chem. 1997 Oct 17;272(42):26145-52
9334180
J Biochem. 2004 Jan;135(1):139-48
14999019
Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):12510-5
14530406
Plant Cell Physiol. 2001 May;42(5):451-61
11382810
Plant Cell. 1995 Jul;7(7):1099-1111
12242400
Proc Natl Acad Sci U S A. 1999 Feb 16;96(4):1480-5
9990049
Biochemistry. 1998 Jun 9;37(23):8282-8
9622480
J Biol Chem. 2009 May 22;284(21):14276-85
19307188
FEMS Yeast Res. 2007 Jan;7(1):102-9
17311588
Plant Physiol. 2001 Jan;125(1):437-46
11154351