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Identification of a functional nuclear export signal in the green fluorescent protein asFP499
Authors:Mustafa Huseyin  Strasser Bernd  Rauth Sabine  Irving Robert A  Wark Kim L
Affiliation:Cooperative Research Centre for Diagnostics at CSIRO Molecular and Health Technologies, 343 Royal Parade, Parkville, Vic. 3052, Australia. huseyinm@hotmail.com
Abstract:The green fluorescent protein (GFP) asFP499 from Anemonia sulcata is a distant homologue of the GFP from Aequorea victoria. We cloned the asFP499 gene into a mammalian expression vector and showed that this protein was expressed in the human lymphoblast cell line Ramos RA1 and in the embryonic kidney 293T cell line (HEK 293T). In HEK 293T cells, asFP499 was localized mainly in the cytoplasm, suggesting that the protein was excluded from the nucleus. We identified (194)LRMEKLNI(201) as a candidate nuclear export signal in asFP499 and mutated the isoleucine at position 201 to an alanine. Unlike the wildtype form, the mutant protein was distributed throughout the cytoplasm and nucleus. This is the first report of a GFP that contains a functional NES.
Keywords:asFP499   GFP   Mammalian cells   Nuclear export signal   Subcellular localization
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