title: Utilizing the Dyn2 dimerization-zipper as a tool to probe NPC structure and function.
abstract: The discovery of dynein light chain 2 (Dyn2) as a member of the nucleoporins in yeast led to a series of applications to study NPC structure and function. Its intriguing ability to act as a hub for the parallel dimerization of two short amino acid sequence motifs (DID) prompted us to utilize it as a tool for probing nucleocytoplasmic transport in vivo. Further, the distinct structure of the Dyn2-DID rod, which is easily visible in the electron microscope, allowed us to develop a precise structural label on proteins or protein complexes. This label was used to identify the position of subunits in NPC subcomplexes or to derive at pseudo-atomic models of single large Nups. The versatility for various applications of the DID-Dyn2 system makes it an attractive molecular tool beyond the nuclear pore and transport field.
authors.affiliation: Biochemie-Zentrum der Universität Heidelberg (BZH), Im Neuenheimer Feld, Heidelberg, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Flemming
authors.affiliation: Biochemie-Zentrum der Universität Heidelberg (BZH), Im Neuenheimer Feld, Heidelberg, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Stelter
authors.affiliation: Biochemie-Zentrum der Universität Heidelberg (BZH), Im Neuenheimer Feld, Heidelberg, Germany.
authors.forename: Ed
authors.initials: E
authors.lastname: Hurt
mesh_terms: D021581:Active Transport, Cell Nucleus; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004398:Dyneins / Q000378:metabolism*; D008969:Molecular Sequence Data; D022022:Nuclear Pore / Q000378:metabolism / Q000648:ultrastructure*; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism / Q000648:ultrastructure*; D011485:Protein Binding; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: D028861:Nuclear Pore Complex Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C500512:intermediate chain 1 protein, S cerevisiae; D004398:Dyneins; C083745:SLC1 protein, S cerevisiae
keywords: DID; Dynein2; Electron microscopy; FG repeats; Label; NPC; Negative stain; Transport channel