Redundant Nuclear Localization Signals Mediate Nuclear Import Of The Aspergillus nidulans Transcription Activator Of Nitrogen Metabolic Genes AreA

Ref ID: 18498

Author:

Cameron C. Hunter,
Kendra S. Siebert,
Damien J. Downes,
Koon Ho Wong,
Sara Lewis,
James A. Fraser,
Michael J. Hynes,
Meryl A. Davis,
Richard B. Todd

Author address:

Department of Plant Pathology, Kansas State University, Manhattan,  KS 66506, USA  
Department of Genetics,
The University of Melbourne, VIC 3010, AUSTRALIA  

Full conference title:

11 th European Conference on Fungal Genetics

Abstract:

The Aspergillus nidulans GATA transcription factor AreA activates transcription of genes for uptake and
metabolism of nitrogen nutrients. AreA accumulates in the nucleus during nitrogen starvation but not in the
presence of nitrogen sources. The AreA protein contains five putative classical SV40 Large T Antigen8208;type nuclear
localization sequences (NLSs) and one putative non8208;canonical bipartite NLS conserved with mammalian GATA4.
We have used two approaches to determine which of these predicted NLSs are functional. First, we fused DNA
sequences encoding the putative AreA NLSs to the Green Fluorescent Protein (GFP) gene and introduced these
constructs into A. nidulans. We determined which of the predicted NLSs are sufficient to direct GFP to the nucleus.
UV8208;fluorescence microscopy showed that the bipartite NLS strongly directs GFP to the nucleus, one of the classical
NLSs weakly directs GFP to the nucleus and the other four classical NLSs collaborate to direct GFP to the nucleus.
Second, we constructed epitope8208;tagged gene replacement areA mutants affected in individual NLSs and
combinations of NLSs to identify by immunofluorescence microscopy sequences required for nuclear localization.
Deletion of all five classical NLSs did not affect utilization of nitrogen sources and did not prevent AreA nuclear
localization. Mutation of the bipartite NLS conferred inability to utilize alternative nitrogen sources but did not
prevent AreA nuclear localization. We determined the effect of this bipartite NLS mutation with combinations of
deletions of the five classical NLSs on nuclear localization. Our results indicate redundancy among the AreA NLSs.

Abstract Number: PR7.8

Conference Year: 2012

Link to conference website: http://www.ecfg.info/images/Abstract_Book_Electronic.pdf

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