G protein betagamma subunits interact with alphabeta- and gamma-tubulin and play a role in microtubule assembly in PC12 cells.
The betagamma subunit of G proteins (Gbetagamma) is known to transfer signals from cell surface receptors to intracellular effector molecules. Recent results suggest that Gbetagamma also interacts with microtubules and is involved in the regulation of the mitotic spindle. In the current study, the anti-microtubular drug nocodazole was employed to investigate the mechanism by which Gbetagamma interacts with tubulin and its possible implications in microtubule assembly in cultured PC12 cells. Nocodazole-induced depolymerization of microtubules drastically inhibited the interaction between Gbetagamma and tubulin. Gbetagamma was preferentially bound to microtubules and treatment with nocodazole suggested that the dissociation of Gbetagamma from microtubules is an early step in the depolymerization process. When microtubules were allowed to recover after removal of nocodazole, the tubulin-Gbetagamma interaction was restored. Unlike Gbetagamma, however, the interaction between tubulin and the alpha subunit of the Gs protein (Gsalpha) was not inhibited by nocodazole, indicating that the inhibition of tubulin-Gbetagamma interactions during microtubule depolymerization is selective. We found that Gbetagamma also interacts with gamma-tubulin, colocalizes with gamma-tubulin in centrosomes, and co-sediments in centrosomal fractions. The interaction between Gbetagamma and gamma-tubulin was unaffected by nocodazole, suggesting that the Gbetagamma-gamma-tubulin interaction is not dependent on assembled microtubules. Taken together, our results suggest that Gbetagamma may play an important and definitive role in microtubule assembly and/or stability. We propose that betagamma-microtubule interaction is an important step for G protein-mediated cell activation. These results may also provide new insights into the mechanism of action of anti-microtubule drugs.
Montoya
Valentina
V
The Department of Biological Sciences, University of Texas El Paso, El Paso, Texas 79968, USA.
Gutierrez
Christina
C
Najera
Omar
O
Leony
Denisse
D
Varela-Ramirez
Armando
A
Popova
Juliana
J
Rasenick
Mark M
MM
Das
Siddhartha
S
Roychowdhury
Sukla
S
D000818:Animals; D002503:Centromere / Q000737:chemistry / Q000378:metabolism; D044387:GTP-Binding Protein beta Subunits / Q000032:analysis / Q000187:drug effects / Q000378:metabolism*; D044388:GTP-Binding Protein gamma Subunits / Q000032:analysis / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008870:Microtubules / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D041681:NIH 3T3 Cells; D015739:Nocodazole / Q000494:pharmacology; D016716:PC12 Cells; D051381:Rats; D014404:Tubulin / Q000032:analysis / Q000378:metabolism*; D050257:Tubulin Modulators / Q000494:pharmacology
D044387:GTP-Binding Protein beta Subunits; D044388:GTP-Binding Protein gamma Subunits; D014404:Tubulin; D050257:Tubulin Modulators; D015739:Nocodazole