From: Nuclear APC
Madame Curie Bioscience Database [Internet].
Austin (TX): Landes Bioscience; 2000-2013.
Copyright © 2000-2013, Landes Bioscience.
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Table 1cEvidence for APC's nuclear export signals (NES)
| Ref | |
|---|---|
| NES1 (aa. 68-77) and NES2 (aa. 165-174) | |
| 1. Each directs nuclear export when fused to GFP-tagged HIV-Rev with inactivated endogenous NES | 8,15 |
| 2. Each directs nuclear export when fused to purified GST and injected into nuclei of living cells | 15 |
| 3. Each can functionally substitute for HIV-Rev NES in in vivo Rev activity assay | 15 |
| 4. Together, mediated Crm1 interaction with a short N-terminal fragment of APC (aa. 1-270) in a mammalian two-hybrid assay | 15 |
| 5. The two NESs were able to facilitate nuclear shuttling of APC fragment (aa 1-270) in a heterokaryon assay | 15 |
| 6. Substitution of alanine for critical leucine residues in each NES inhibited the activities measured in all assays listed (1-5) | 8,15 |
| 7. Mutation of either NES in the context of full-length exogenous APC resulted in shift of APC from predominantly cytoplasmic to more nuclear than cytoplasmic | 8,15 |
| 8. Purified monomeric fragment of APC (aa. 129-250) containing only NES2 bound the nuclear exportin protein Crm1 in vitro | 16 |
| NES-R3 (aa. 1506-1511), NES-R4 (aa. 1657-1662) and NES-R7 (aa. 2027-2032) | |
| 1. Each directs nuclear export when fused to GFP-tagged N-terminal fragment of Drosophila APC2, dAPC2 (aa. 114-410) | 19 |
| 2. dAPC2 (aa. 460-1067) containing NES-R3 and -R4 and fused with GFP was cytoplasmic when expressed in COS cells, but more nuclear when cells were treated with LMB | 19 |
| 3. APC (aa. 1379-2080) containing NES-R3, -R4 and R7 behaved similarly | 19 |
| 4. APC (aa. 1379-2080) showed a loss of 50% nuclear fluorescence after 20 seconds of sequential photobleaching of the cytoplasm | 20 |