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PMID:20126275
| Citation |
Kaczmarczyk, SJ, Sitaraman, K, Hill, T, Hartley, JL and Chatterjee, DK (2010) Tus, an E. coli protein, contains mammalian nuclear targeting and exporting signals. PLoS ONE 5:e8889 |
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| Abstract |
Shuttling of proteins between nucleus and cytoplasm in mammalian cells is facilitated by the presence of nuclear localization signals (NLS) and nuclear export signals (NES), respectively. However, we have found that Tus, an E. coli replication fork arresting protein, contains separate sequences that function efficiently as NLS and NES in mammalian cell lines, as judged by cellular location of GFP-fusion proteins. The NLS was localized to a short stretch of 9 amino acids in the carboxy-terminus of Tus protein. Alterations of any of these basic amino acids almost completely abolished the nuclear targeting. The NES comprises a cluster of leucine/hydrophobic residues located within 21 amino acids at the amino terminus of Tus. Finally, we have shown that purified GFP-Tus fusion protein or GFP-Tus NLS fusion protein, when added to the culture media, was internalized very efficiently into mammalian cells. Thus, Tus is perhaps the first reported bacterial protein to possess both NLS and NES, and has the capability to transduce protein into mammalian cells. |
| Links |
PubMed PMC2811178 Online version:10.1371/journal.pone.0008889 |
| Keywords |
Amino Acid Sequence; Base Sequence; Cell Line, Tumor; Cell Nucleus/metabolism; DNA Primers; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/metabolism; Humans; Male; Molecular Sequence Data; Nuclear Export Signals |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0006607: NLS-bearing substrate import into nucleus |
IMP: Inferred from Mutant Phenotype: |
P |
A series of N- and C- terminal deletion mutants of Tus were constructed which were fused to GFP to determine the subcellular distribution of green fluorescence in the transduced cells. Figure 2A shows that the fusion of amino acids 218 to 309 of Tus to the C-terminus of GFP restored nuclear targeting, defining the NLS sequence of Tus. Further depicted in Figure 3A, alteration of any one of the basic amino acids 218-309 resulted in pronounced perturbation of nuclear targeting of the fusion proteins. |
complete |
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See also
References
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