Multiple nuclear localization signals mediate nuclear localization of the GATA transcription factor AreA

Hunter, Cameron C., Siebert, Kendra S., Downes, Damien J., Wong, Koon Ho, Kreutzberger, Sara D., Fraser, James A., Clarke, David F., Hynes, Michael J., Davis, Meryl A. and Todd, Richard B. (2014) Multiple nuclear localization signals mediate nuclear localization of the GATA transcription factor AreA. Eukaryotic Cell, 13 4: 527-538. doi:10.1128/EC.00040-14


Author Hunter, Cameron C.
Siebert, Kendra S.
Downes, Damien J.
Wong, Koon Ho
Kreutzberger, Sara D.
Fraser, James A.
Clarke, David F.
Hynes, Michael J.
Davis, Meryl A.
Todd, Richard B.
Title Multiple nuclear localization signals mediate nuclear localization of the GATA transcription factor AreA
Journal name Eukaryotic Cell   Check publisher's open access policy
ISSN 1535-9778
1535-9786
Publication date 2014-04-01
Year available 2014
Sub-type Article (original research)
DOI 10.1128/EC.00040-14
Open Access Status Not Open Access
Volume 13
Issue 4
Start page 527
End page 538
Total pages 12
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Language eng
Formatted abstract
The Aspergillus nidulans GATA transcription factor AreA activates transcription of nitrogen metabolic genes in response to nitrogen limitation and is known to accumulate in the nucleus during nitrogen starvation. Sequence analysis of AreA revealed multiple nuclear localization signals (NLSs), five putative classical NLSs conserved in fungal AreA orthologs but not in the Saccharomyces cerevisiae functional orthologs Gln3p and Gat1p, and one putative noncanonical RRX33RXR bipartite NLS within the DNA-binding domain. In order to identify the functional NLSs in AreA, we constructed areA mutants with mutations in individual putative NLSs or combinations of putative NLSs and strains expressing green fluorescent protein (GFP)-AreA NLS fusion genes. Deletion of all five classical NLSs individually or collectively did not affect utilization of nitrogen sources or AreAdependent gene expression and did not prevent AreA nuclear localization. Mutation of the bipartite NLS conferred the inability to utilize alternative nitrogen sources and abolished AreA-dependent gene expression likely due to effects on DNA binding but did not prevent AreA nuclear localization. Mutation of all six NLSs simultaneously prevented AreA nuclear accumulation. The bipartite NLS alone strongly directed GFP to the nucleus, whereas the classical NLSs collaborated to direct GFP to the nucleus. Therefore, AreA contains multiple conserved NLSs, which show redundancy and together function to mediate nuclear import. The noncanonical bipartite NLS is conserved in GATA factors from Aspergillus, yeast, and mammals, indicating an ancient origin.
Keyword Microbiology
Mycology
Microbiology
Mycology
MICROBIOLOGY
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID DP0558002
P20RR016475
EPS-0903806
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Non HERDC
School of Chemistry and Molecular Biosciences
 
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