Role of different domains in the self-association of rat nucleoporin p62.
We have expressed rat nucleoporin p62 cDNA in Escherichia coli to obtain material for structural and self-association studies. Electron microscopy and circular dichroism spectroscopy are consistent with a rod-shaped molecule with an alpha-helical coiled-coil domain at its C terminus and a cross-beta structure at its N terminus, separated by a threonine-rich linker, which has a less-defined secondary structure. Electron microscopy and the solubility properties of fragments produced using thrombin and CNBr digestion indicate that p62 molecules associate to form linear chains and that a small region near the C terminus is an important determinant of assembly. This association may have important consequences for pore structure and function; for example, one way p62 could associate would be to form rings in nuclear pores that could function like barrel hoops.
Buss
F
F
MRC Laboratory of Molecular Biology, Cambridge, UK.
Kent
H
H
Stewart
M
M
Bailer
S M
SM
Hanover
J A
JA
D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics; D008854:Microscopy, Electron; D008956:Models, Chemical; D008969:Molecular Sequence Data; D015394:Molecular Structure; D028861:Nuclear Pore Complex Proteins; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012995:Solubility
D018076:DNA, Complementary; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D028861:Nuclear Pore Complex Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C049312:nuclear pore protein p62