Early recruitment of AU-rich element-containing mRNAs determines their cytosolic fate during iron deficiency.
The yeast Cth2 protein is a CX(8)CX(5)CX(3)H tandem zinc finger protein that binds AU-rich element (ARE)-containing transcripts to enhance their decay in response to iron (Fe) deficiency. Mammalian members of this family of proteins are known to undergo nucleocytoplasmic shuttling, but little is known about the role of shuttling in the mechanism of ARE-dependent mRNA decay. Here we demonstrate that, like its mammalian homologues, Cth2 is a nucleocytoplasmic shuttling protein whose nuclear export depends on mRNA transport to the cytosol. The nuclear import information of Cth2 is contained within its tandem zinc finger domain, but it is independent of mRNA-binding function. Moreover, we also demonstrate that nucleocytoplasmic shuttling of Cth2 requires active transcription and that disruption of shuttling leads to defects in Cth2 function in mRNA decay under Fe deficiency. Taken together, our data suggest that under conditions of Fe deficiency Cth2 travels into the nucleus to recruit target mRNAs, perhaps cotranscriptionally, that are destined for cytosolic degradation as part of the mechanism of adaptation to growth under Fe limitation. These data also suggest an important role for nucleocytoplasmic shuttling in this conserved family of proteins in the mechanism of ARE-mediated mRNA decay.
Department of Pharmacology and Cancer Biology, Duke University Medical Center, Research Drive-LSRC-C134, Durham, NC 27710, USA.
Sandra V
SV
Vergara
Sergi
S
Puig
Dennis J
DJ
Thiele
D021581:Active Transport, Cell Nucleus; D002467:Cell Nucleus / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007501:Iron / Q000378:metabolism; D000090463:Iron Deficiencies; D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D020871:RNA Stability; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D038621:Regulatory Sequences, Ribonucleic Acid / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014158:Transcription, Genetic
D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D038621:Regulatory Sequences, Ribonucleic Acid; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; D007501:Iron