Cytoplasmic sequestration of rel proteins by IkappaBalpha requires CRM1-dependent nuclear export.
Rel and IkappaB protein families form a complex cellular regulatory network. A major regulatory function of IkappaB proteins is to retain Rel proteins in the cell cytoplasm. In addition, IkappaB proteins have also been postulated to serve nuclear functions. These include the maintenance of inducible NF-kappaB-dependent gene transcription, as well as termination of inducible transcription. We show that IkappaBalpha shuttles between the nucleus and the cytoplasm, utilizing the nuclear export receptor CRM1. A CRM1-binding export sequence was identified in the N-terminal domain of IkappaBalpha but not in that of IkappaBbeta or IkappaBepsilon. By reconstituting major aspects of NF-kappaB-IkappaB sequestration in yeast, we demonstrate that cytoplasmic retention of p65 (also called RelA) by IkappaBalpha requires Crm1p-dependent nuclear export. In mammalian cells, inhibition of CRM1 by leptomycin B resulted in nuclear localization of cotransfected p65 and IkappaBalpha in COS cells and enhanced nuclear relocation of endogenous p65 in T cells. These observations suggest that the main function of IkappaBalpha is that of a nuclear export chaperone rather than a cytoplasmic tether. We propose that the nucleus is the major site of p65-IkappaBalpha association, from where these complexes must be exported in order to create the cytoplasmic pool.
Tam
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Rosenstiel Basic Medical Sciences Research Center, Waltham, Massachusetts 02454, USA.
Lee
L H
LH
Davis
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Sen
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D000818:Animals; D001692:Biological Transport; D019556:COS Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D020573:Genes, rel; D020672:I-kappa B Proteins / Q000378:metabolism*; D028884:Karyopherins; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D020572:Oncogene Proteins v-rel / Q000235:genetics / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear; D012441:Saccharomyces cerevisiae; D000097606:Exportin 1 Protein
D002352:Carrier Proteins; D020672:I-kappa B Proteins; D028884:Karyopherins; D009687:Nuclear Proteins; D020572:Oncogene Proteins v-rel; D018160:Receptors, Cytoplasmic and Nuclear; D000097606:Exportin 1 Protein