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| Sec13 shuttles between the nucleus and the cytoplasm and stably interacts with Nup96 at the nuclear pore complex |
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| Author(s): Enninga J, Levay A, Fontoura BMA |
| Source: MOLECULAR AND CELLULAR BIOLOGY Volume: 23 Issue: 20 Pages: 7271-7284 Published: OCT 2003 |
| Times Cited: 34 References: 56 |
| Abstract: Sec13 is a constituent of the endoplasmic reticulum and the nuclear pore complex (NPC). At the endoplasmic reticulum, Sec13 is involved in the biogenesis of COPII-coated vesicles, whereas at the NPC its function is unknown. We show here, by yeast two-hybrid screenings and biochemical assays, that a region at the amino terminus of the human nuclear pore complex protein Nup96 interacts with the WD (Trp-Asp) repeat region of human Sec13. By using immunofluorescence and confocal and immunoelectron microscopy, we found that in interphase, Sec13 and Nup96 are localized at both sides of the NPC in addition to other intracellular sites. In mitosis, Sec13 was found dispersed throughout the cell, whereas a pool of Nup96 colocalized with the spindle apparatus. Photobleaching experiments showed that Sec13 shuttles between intranuclear sites and the cytoplasm, and a fraction of Sec13 is stably associated with NPCs. Cotransfection of Sec13 and the Sec13 binding site of Nup96 decreased the mobile pool of Sec][3, demonstrating the interaction of Sec13 and Nup96 in vivo. Targeting studies showed that Sec13 is actively transported into the nucleus and contains a nuclear localization signal. These results indicate that Sec13 stably interacts with Nup96 at the NPC during interphase and that the shuttling of Sec13 between the nucleus and the cytoplasm may couple and regulate functions between these two compartments. |
| Document Type: Article |
| Language: English |
| Reprint Address: Fontoura, BMA (reprint author), Univ Miami, Sch Med, Dept Mol & Cellular Pharmacol, 1600 NW 10th Ave, Miami, FL 33136 USA |
Addresses:
1. Univ Miami, Sch Med, Dept Mol & Cellular Pharmacol, Miami, FL 33136 USA
2. Univ Miami, Sch Med, Sylvester Canc Ctr, Miami, FL 33136 USA |
| Publisher: AMER SOC MICROBIOLOGY, 1752 N ST NW, WASHINGTON, DC 20036-2904 USA |
| Subject Category: Biochemistry & Molecular Biology; Cell Biology |
| IDS Number: 729CT |
| ISSN: 0270-7306 |
| DOI: 10.1128/MCB.23.20.7271-7284.2003 |
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