Phosphorylation by cdc2-CyclinB1 Kinase Releases Cytoplasmic Dynein from Membranes

Abstract

Movement of various cargoes toward microtubule minus ends is driven by the microtubule motor cytoplasmic dynein (CD). Many cargoes are motile only during certain cell cycle phases, suggesting that CD function may be under cell cycle control. Phosphorylation of the CD light intermediate chain (DLIC) has been suggested to play a crucial role in modulating CD function during the Xenopus embryonic cell cycle, where CD-driven organelle movement is active in interphase but greatly reduced in metaphase. This down-regulation correlates with hyperphosphorylation of DLIC and release of CD from the membrane. Here we investigate the role of the key mitotic kinase, cdc2-cyclinB1, in this process. We show that DLIC within the native Xenopus CD complex is an excellent substrate for purified Xenopus cdc2-glutathione S-transferase (GST) cyclinB1 (cdc2-GSTcyclinB1) kinase. Mass spectrometry of native DLIC revealed that a conserved cdc2 site (Ser-197) previously implicated in the metaphase modulation of CD remains phosphorylated in interphase and so is unlikely to be the key regulatory site. We also demonstrate that incubating interphase membranes with cdc2-GSTcyclinB1 kinase results in substantial release of CD from the membrane. These data suggest that phosphorylation of DLIC by cdc2 kinase leads directly to the loss of membrane-associated CD and an inhibition of organelle movement.

Rab11-FIP3 links the Rab11 GTPase and cytoplasmic dynein to mediate transport to the endosomal-recycling compartment

“…dynein and lead to inhibition of organelle movement (Addinall et al, 2001;Dell et al, 2000;Niclas et al, 1996;Sivaram et al, 2009).…”

Functional coupling of microtubules to membranes – implications for membrane structure and dynamics

“…An example of how this interaction can be dynamically affected is the regulation of the association between motors and their cargoes by protein phosphorylation (Yeh et al, 2006;Guillaud et al, 2008). Similarly, phosphorylation of the dynein LIC1 by cyclindependent kinase 1 (CDK1) leads to the dissociation of the light chain from the membrane (Niclas et al, 1996;Addinall et al, 2001). …”

Biased inheritance of mitochondria during asymmetric cell division in the mouse oocyte

“…Also necessary is progression through meiosis I. A direct role for the decrease in Cdk1 activity on motors (Addinall et al, 2001;Niclas et al, 1996) or cytoskeletal organization (Field et al, 2011) is one possible mechanism for withdrawing mitochondria to the oocyte-directed pole of the spindle. Alternatively, forces associated with cortical contraction and associated cytoplasmic reorganization (Brunet and Verlhac, 2011) may repel mitochondria from the polarized cortex destined to become the polar body.…”