Transcriptional regulation of the Zrg17 zinc transporter of the yeast secretory pathway.
The Msc2 and Zrg17 proteins of Saccharomyces cerevisiae are members of the cation diffusion facilitator family of zinc transporters. These proteins form heteromeric complexes that transport zinc into the ER (endoplasmic reticulum). Previous studies suggested that the ZRG17 gene is regulated in response to zinc status by the Zap1 transcription factor. Zap1 activates the expression of many genes in zinc-deficient cells. In the present study, we assessed whether ZRG17 is a direct Zap1 target gene. We showed that ZRG17 mRNA levels were elevated in zinc-limited cells in a Zap1-dependent manner and were also elevated in zinc-replete cells expressing a constitutively active allele of Zap1. Furthermore, Zrg17 protein levels correlated closely with mRNA levels. A candidate Zap1-binding site [ZRE (zinc-responsive element)] in the ZRG17 promoter was required for this induction. Using electrophoretic mobility-shift assays and chromatin immunoprecipitation, we demonstrated that Zap1 binds specifically to the ZRG17 ZRE both in vitro and in vivo. By using a chromosomal ZRG17 mutant with a non-functional ZRE, we found that Zap1 induction of ZRG17 is required for ER function as indicated by elevated ER stress under zinc-limited conditions. Together, these results establish that ZRG17 is a direct Zap1 target gene and its regulation has biological importance in maintaining ER function.
Department of Nutritional Sciences, University of Wisconsin-Madison, 1415 Linden Drive, Madison, WI 53706, USA.
Yi-Hsuan
YH
Wu
Avery G
AG
Frey
David J
DJ
Eide
D000483:Alleles; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006706:Homeostasis; D050296:Microbial Viability; D009154:Mutation; D009994:Osmolar Concentration; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D018085:Regulon; D020218:Response Elements / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D055571:Secretory Pathway; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D015032:Zinc / Q000378:metabolism*
D027682:Cation Transport Proteins; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C103264:ZAP1 protein, S cerevisiae; C558193:zrg17 protein, S cerevisiae; D015032:Zinc
Methods Enzymol. 1987;154:164-75
3323810
BMC Genomics. 2008 Aug 01;9:370
18673560
J Biol Chem. 2002 Sep 13;277(37):33749-57
12095998
Gene. 1986;45(3):299-310
3026915
EMBO J. 2006 Dec 13;25(24):5726-34
17139254
J Biol Chem. 2009 Oct 2;284(40):27544-56
19656949
Nat Biotechnol. 2008 Dec;26(12):1339-40
19060867
J Cell Biol. 2004 Aug 2;166(3):325-35
15277543
Genetics. 2000 Sep;156(1):45-58
10978274
Nat Struct Mol Biol. 2009 Oct;16(10):1063-7
19749753
J Biol Chem. 2000 May 26;275(21):16160-6
10747942
Science. 2007 Sep 21;317(5845):1746-8
17717154
PLoS One. 2008;3(11):e3642
18985159
J Biol Chem. 2003 Apr 25;278(17):15065-72
12556516
J Biol Chem. 2003 Feb 7;278(6):4096-102
12446736
J Biol Chem. 2005 Aug 5;280(31):28811-8
15961382
Yeast. 1994 Jan;10(1):105-12
7515538
J Biol Chem. 1996 Sep 20;271(38):23203-10
8798516
EMBO J. 2004 Mar 10;23(5):1123-32
14976557
EMBO J. 2000 Jun 15;19(12):2845-55
10856230
Mol Biol Cell. 1997 Oct;8(10):1845-62
9348528
Cell. 2000 Apr 28;101(3):249-58
10847680
Proc Natl Acad Sci U S A. 2007 Apr 3;104(14):5812-7
17392431
Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):7957-62
10884426
J Mol Biol. 2006 Apr 7;357(4):1167-83
16483601
Methods Enzymol. 1983;101:181-91
6310321
Genome Res. 2009 Apr;19(4):556-66
19158363
J Biol Chem. 1998 Oct 30;273(44):28713-20
9786867
Genes Dev. 1992 Jan;6(1):93-104
1730413
Nucleic Acids Res. 2005 May 19;33(9):2838-51
15905473
Am J Hum Genet. 2008 Jun;82(6):1290-305
18513683
J Biol Chem. 2007 Apr 6;282(14):10423-31
17234632
J Biol Chem. 1998 Oct 30;273(44):28617-24
9786854
J Cell Biol. 2007 May 21;177(4):637-45
17502426
EMBO J. 2000 Jul 17;19(14):3704-13
10899124
J Biol Chem. 2009 Jul 10;284(28):18565-9
19363031
Nat Biotechnol. 2001 Aug;19(8):773-6
11479573
J Biol Chem. 2009 Nov 6;284(45):30798-806
19759014
Biochem Pharmacol. 2004 Aug 15;68(4):699-709
15276077
Proc Natl Acad Sci U S A. 1996 Mar 19;93(6):2454-8
8637895
Mol Microbiol. 2005 Aug;57(3):834-46
16045625
J Biol Chem. 2005 Sep 2;280(35):30956-62
15994300
Mol Cell Biol. 1997 Sep;17(9):5044-52
9271382
J Biol Chem. 2009 Jun 26;284(26):17677-86
19366695
J Biol Chem. 2006 Jun 30;281(26):17743-50
16636052