MZT1 regulates microtubule nucleation by linking γTuRC assembly to adapter-mediated targeting and activation

Regulation of the γ-tubulin ring complex (γTuRC) through targeting and activation restricts nucleation of microtubules to microtubule-organizing centers (MTOCs), aiding in the assembly of ordered microtubule arrays. However, the mechanistic basis of this important regulation remains poorly understood. Here, we show that, in human cells, γTuRC integrity, determined by the presence of γ-tubulin complex proteins (GCPs; also known as TUBGCPs) 2–6, is a prerequisite for interaction with the targeting factor NEDD1, impacting on essentially all γ-tubulin-dependent functions. Recognition of γTuRC integrity is mediated by MZT1, which binds not only to the GCP3 subunit as previously shown, but cooperatively also to other GCPs through a conserved hydrophobic motif present in the N-termini of GCP2, GCP3, GCP5 and GCP6. MZT1 knockdown causes severe cellular defects under conditions that leave γTuRC intact, suggesting that the essential function of MZT1 is not in γTuRC assembly. Instead, MZT1 specifically binds fully assembled γTuRC to enable interaction with NEDD1 for targeting, and with the CM1 domain of CDK5RAP2 for stimulating nucleation activity. Thus, MZT1 is a ‘priming factor’ for γTuRC that allows spatial regulation of nucleation.
This study was supported by grants from the Ministerio de Economıa y ́ Competitividad (MINECO) (BFU2009-08522, BFU2012-33960 and BFU2015- 69275-P to J.L. and BFU2014-58422-P to J.R.), and IRB Barcelona intramural funds. J.L. acknowledges additional support from the Ramón y Cajal Programme (RYC-2010-07108 MINECO, Spain). R.R.C. was supported by a PhD fellowship from the Consejo Nacional de Ciencia y Tecnologıa (Mexican National Council for ́ Science and Technology; CONACYT) (CVU 269229).