Identification of a functional nuclear export signal in the green fluorescent protein asFP499.
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.
Mustafa
Huseyin
H
Cooperative Research Centre for Diagnostics at CSIRO Molecular and Health Technologies, 343 Royal Parade, Parkville, Vic. 3052, Australia. huseyinm@hotmail.com
Strasser
Bernd
B
Rauth
Sabine
S
Irving
Robert A
RA
Wark
Kim L
KL
D000595:Amino Acid Sequence; D002460:Cell Line; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism*; D008969:Molecular Sequence Data; D049790:Nuclear Export Signals / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism*
D049790:Nuclear Export Signals; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins