Molecular characterization of a TIA-1-like RNA-binding protein in cells derived from the fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae).
A complementary DNA encoding a TIA-1-type RNA-binding protein (SfTRN-1) was isolated from cultured cells of the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), to characterize its function. The deduced 388-amino acid sequence of SfTRN-1, which possessed three RNA recognition motifs (RRMs) followed by a C-terminal auxiliary domain, showed significant homology with mammalian TIA-1/TIAR and silkworm BmTRN-1, factors important in the metabolism of transcripts. It was found that inhibition of SfTRN-1 gene expression by a transfected oligonucleotide encoding the antisense sequence led to a marked increase in the production of a reporter protein and the amount of reporter transcript in the cultured cells. In addition, overexpression of the recombinant full-length SfTRN-1 open reading frame in the cultured cells led to a decrease in reporter protein production, but the truncated RRM1-3 domain lacking the C-terminal auxiliary domain lost its activity. Analysis using a GFP-fused recombinant protein revealed that, unlike mammalian TIA-1/TIAR, SfTRN-1, most likely shuttling between the nucleus and cytoplasm, had the characteristic of being largely distributed in the cytoplasm, where it perhaps acts to reduce the amount of transcripts, and that RRM1 and RRM3 were related to its nuclear accumulation, but RRM2 to its nuclear export. Furthermore, the posterior half of the auxiliary domain was also found to be related to its nuclear export. This study indicates that respective RRM subdomains of SfTRN-1 play distinct roles important to its subcellular distribution, and it identified unique systems for the distribution and functional regulation of the TIA-1 family in insect cells, ones which are clearly different from those in mammalian cells.
Muto
Sayaka
S
Graduate School of Science and Technology, Kyoto Institute of Technology, Sakyo, Kyoto, Japan.
Tanabe
Toru
T
Matsumoto
Emi
E
Mori
Hajime
H
Kotani
Eiji
E
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D042541:Intracellular Space / Q000378:metabolism; D008969:Molecular Sequence Data; D012323:RNA Processing, Post-Transcriptional; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D018411:Spodoptera / Q000166:cytology*; D013312:Stress, Physiological
D016601:RNA-Binding Proteins