Functional characterization of nuclear localization signals in theAspergillus nidulans transcription activator of nitrogen metabolic genes AreA.

Ref ID: 18338

Author:

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

Author address:

Department of Plant Pathology, Kansas State University, Manhattan, KS 66506, USA. Department of Genetics, The University of
Melbourne, VIC 3010, AUSTRALIA. Email: rbtodd@k-state.edu

Full conference title:

Asperfest 8

Abstract:

The Aspergillus nidulans GATADNA-binding transcription factor AreAactivates 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 Antigen-type nuclear localization sequences (NLSs) and one putative non-canonical bipartite NLS conserved with mammalian GATA4.
We are using two approachesto determinewhich of the putative NLSs are functional. First, we constructed epitope-tagged gene replacement areA mutants
affected in the NLSs to identify sequences required for nuclear localization. Immunofluorescence microscopy experiments show that at least one of the
classical NLSs contributes to nuclear import. However, deletion of all five classical NLSs does not affect utilization of nitrogen sources and does not
prevent AreA nuclear localization. Mutation of the bipartite NLS confers inability to utilize alternative nitrogen sources. We are determining the effect
of this bipartite NLS mutation individually and in combination with deletion of the five classical NLSs on nuclear localization. Second, we fused DNA
sequences encoding the putative AreA NLSs to the Green Fluorescent Protein (GFP) gene and introduced these constructs into A. nidulans to determine
which of the predicted NLSs are sufficient to direct GFP to the nucleus. We will use UV-fluorescence microscopy to determine the subcellular location
of the GFP-NLS fusion protein in the transformants.

Abstract Number: 16)

Conference Year: 2011

Link to conference website: NULL

New link: NULL

Conference abstracts, posters & presentations

Showing 10 posts of 17325 posts found.

  • Title

    Author

    Year

    Number

    Poster