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Purification of NSP1 reveals complex formation with ‘GLFG’ nucleoporins and a novel nuclear pore protein NIC96.
Author(s) -
Grandi P.,
Doye V.,
Hurt E.C.
Publication year - 1993
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1993.tb05975.x
Subject(s) - nucleoporin , nuclear pore , biology , domain (mathematical analysis) , nuclear protein , gene , microbiology and biotechnology , genetics , nucleus , transcription factor , mathematical analysis , mathematics
The essential C‐terminal domain of NSP1 mediates assembly into the nuclear pore complex (NPC). To identify components which interact physically with this yeast nucleoporin, the tagged C‐terminal domain of NSP1 (ProtA‐NSP1) was isolated by affinity chromatography under non‐denaturing conditions. The purified complex contains ProtA‐NSP1, two previously identified ‘GLFG’ nucleoporins, NUP49 (NSP49) and p54 and a novel protein designated NIC96 (for Nucleoporin‐Interacting Component of 96 kDa). Conversely, affinity purification of tagged NSP49 enriches for NSP1, the p54 and the NIC96 component. The NIC96 gene was cloned; it encodes a novel 839 amino acid protein essential for cell growth. By immunofluorescence, protein A‐tagged NIC96 exhibits a punctate nuclear membrane staining indicative of nuclear pore location. Therefore, affinity purification of tagged nucleoporins has allowed the definition of a subcomplex of the NPC and analysis of physical interactions between nuclear pore proteins.