A direct interaction with NEDD1 regulates gamma-tubulin recruitment to the centrosome.
The centrosome is the primary microtubule organizing centre of the cell. gamma-tubulin is a core component of the centrosome and is required for microtubule nucleation and centrosome function. The recruitment of gamma-tubulin to centrosomes is mediated by its interaction with NEDD1, a WD40-repeat containing protein. Here we demonstrate that NEDD1 is likely to be oligomeric in vivo and binds directly to gamma-tubulin through a small region of just 62 residues at the carboxyl-terminus of the protein. This carboxyl-terminal domain that binds gamma-tubulin has a helical structure and is a stable tetramer in solution. Mutation of residues in NEDD1 that disrupt binding to gamma-tubulin result in a mis-localization of gamma-tubulin away from the centrosome. Hence, this study defines the binding site on NEDD1 that is required for its interaction with gamma-tubulin, and shows that this interaction is required for the correct localization of gamma-tubulin.
Manning
Jantina A
JA
Centre for Cancer Biology, SA Pathology, Adelaide, South Australia, Australia.
Shalini
Sonia
S
Risk
Joanna M
JM
Day
Catherine L
CL
Kumar
Sharad
S
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D018385:Centrosome / Q000378:metabolism*; D002942:Circular Dichroism; D006801:Humans; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009154:Mutation; D041681:NIH 3T3 Cells; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012542:Scattering, Radiation; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*
D008869:Microtubule-Associated Proteins; C509189:NEDD1 protein, human; D014404:Tubulin; C086844:Nedd1 protein, mouse