Abstract

Interphase microtubule arrays play critical roles in a variety of cellular functions, including the spatial organization and distribution of the endoplasmic reticulum (ER). However, the role of the ER in regulation of microtubule assembly remains poorly characterized. Here, we identify Erg28, a conserved transmembrane protein localized to the ER, as a key factor that inhibits microtubule assembly. Biochemical analyses demonstrate that Erg28 physically interacts with the microtubule assembly-promoting factors—the Mto1-Mto2 complex and the γ-tubulin small complex (γ-TuSC)—and significantly attenuates the binding of γ-TuSC to the Mto1-Mto2 complex. Additionally, microscopic analyses show that Erg28 inhibits microtubule assembly mediated by Mto1-Mto2 complex and γ-TuSC in vitro . The cytosolic N-terminal region of Erg28 is indispensable for its inhibitory activity. Moreover, erg28 deletion leads to excessive microtubule assembly, causing nuclear shape deformation. These findings provide insights into the regulatory mechanism by which the ER influences microtubule cytoskeleton organization.