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ALADIN^{I482S}$Causes Selective Failure of Nuclear Protein Import and Hypersensitivity to Oxidative Stress in Triple A Syndrome
Triple A syndrome is an autosomal recessive neuroendocrinological disease caused by mutations in a gene that encodes 546 amino acid residues. The encoded protein is the nucleoporin ALADIN, a component of nuclear pore complex (NPC). We identified a mutant $ALADIN^{1482S}$ that fails to target NPC and...
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Published in: | Proceedings of the National Academy of Sciences - PNAS 2006-02, Vol.103 (7), p.2298-2303 |
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Main Authors: | , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Online Access: | Get full text |
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Summary: | Triple A syndrome is an autosomal recessive neuroendocrinological disease caused by mutations in a gene that encodes 546 amino acid residues. The encoded protein is the nucleoporin ALADIN, a component of nuclear pore complex (NPC). We identified a mutant $ALADIN^{1482S}$ that fails to target NPC and investigated the consequences of mistargeting using cultured fibroblasts (1482Sf) from a patient with triple A syndrome. $ALADIN^{1482S}$ affected a karyopherin-α/β-mediated import pathway and decreased nuclear accumulations of aprataxin (APTX), a repair protein for DNA singlestrand breaks (SSBs), and of DNA ligase I in 1482Sf. This decrease was restored by wild-type ALADIN. $ALADIN^{1482S}$ had no effect on imports of M9/kap-β2, BIB/kap-β3, histone H1/importin 7, the ubiquitin conjugating enzyme UbcM2/importin 11, or the spliceosome protein U1A, indicating that $ALADIN^{1482S}$ selectively impaired transport of discrete import complexes through NPC. Cell survival assay showed hypersensitivity of 1482Sf to L-buthionine-(S,R)sulfoximine (BSO), a glutathione-depleting agent. BSO decreased nuclear APTX and ligase I levels in 1482Sf and normal control fibroblasts, but increased SSBs only in 1482Sf. These observations implied that 1482Sf are hypersensitive to BSO and no longer sufficiently repair SSBs. Consistent with this notion, 1482Sf transfected with both APTX and ligase I had increased resistance to BSO, whereas 1482Sf transfected with LacZ vector remained hypersensitive to BSO. We propose that oxidative stress aggravates nuclear import failure, which is already compromised in patient cells. Consequent DNA damage, beyond the limited capacity of DNA repair proteins, i.e., APTX and ligase I, may participate in triggering cell death. |
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ISSN: | 0027-8424 |
DOI: | 10.1073/pnas.0505598103 |