The CRR1 nutritional copper sensor in Chlamydomonas contains two distinct metal-responsive domains.
Copper response regulator 1 (CRR1), an SBP-domain transcription factor, is a global regulator of nutritional copper signaling in Chlamydomonas reinhardtii and activates genes necessary during periods of copper deficiency. We localized Chlamydomonas CRR1 to the nucleus in mustard (Sinapis alba) seedlings, a location consistent with its function as a transcription factor. The Zn binding SBP domain of CRR1 binds copper ions in vitro. Cu(I) can replace Zn(II), but the Cu(II) form is unstable. The DNA binding activity is inhibited in vitro by Cu(II) or Hg(II) ions, which also prevent activation of transcription in vivo, but not by Co(II) or Ni(II), which have no effect in vivo. Copper inhibition of DNA binding is reduced by mutation of a conserved His residue. These results implicate the SBP domain in copper sensing. Deletion of a C-terminal metallothionein-like Cys-rich domain impacted neither nutritional copper signaling nor the effect of mercuric supplementation, but rendered CRR1 insensitive to hypoxia and to nickel supplementation, which normally activate the copper deficiency regulon in wild-type cells. Strains carrying the crr1-ΔCys allele upregulate ZRT genes and hyperaccumulate Zn(II), suggesting that the effect of nickel ions may be revealing a role for the C-terminal domain of CRR1 in zinc homeostasis in Chlamydomonas.
Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095-1569, USA.
Frederik
F
Sommer
Janette
J
Kropat
Davin
D
Malasarn
Nicholas E
NE
Grossoehme
Xiaohua
X
Chen
David P
DP
Giedroc
Sabeeha S
SS
Merchant
D000595:Amino Acid Sequence; D016825:Chlamydomonas reinhardtii / Q000378:metabolism*; D016368:Frameshift Mutation; D006706:Homeostasis; D008670:Metals / Q000494:pharmacology*; D008969:Molecular Sequence Data; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D008670:Metals; D014157:Transcription Factors