Tubulin, Gq, and phosphatidylinositol 4,5-bisphosphate interact to regulate phospholipase Cbeta1 signaling.
The cytoskeletal protein, tubulin, has been shown to regulate adenylyl cyclase activity through its interaction with the specific G protein alpha subunits, Galphas or Galphai1. Tubulin activates these G proteins by transferring GTP and stabilizing the active nucleotide-bound Galpha conformation. To study the possibility of tubulin involvement in Galphaq-mediated phospholipase Cbeta1 (PLCbeta1) signaling, the m1 muscarinic receptor, Galphaq, and PLCbeta1 were expressed in Sf9 cells. A unique ability of tubulin to regulate PLCbeta1 was observed. Low concentrations of tubulin, with guanine nucleotide bound, activated PLCbeta1, whereas higher concentrations inhibited the enzyme. Interaction of tubulin with both Galphaq and PLCbeta1, accompanied by guanine nucleotide transfer from tubulin to Galphaq, is suggested as a mechanism for the enzyme activation. The PLCbeta1 substrate, phosphatidylinositol 4,5-bisphosphate, bound to tubulin and prevented microtubule assembly. This observation suggested a mechanism for the inhibition of PLCbeta1 by tubulin, since high tubulin concentrations might prevent the access of PLCbeta1 to its substrate. Activation of m1 muscarinic receptors by carbachol relaxed this inhibition, probably by increasing the affinity of Galphaq for tubulin. Involvement of tubulin in the articulation between PLCbeta1 signaling and microtubule assembly might prove important for the intracellular governing of a broad range of cellular events.
Popova
J S
JS
Department of Physiology and Biophysics and the Committee on Neuroscience, University of Illinois College of Medicine, Chicago, Illinois 60612, USA.
Garrison
J C
JC
Rhee
S G
SG
Rasenick
M M
MM
D000818:Animals; D002462:Cell Membrane / Q000502:physiology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006165:Guanylyl Imidodiphosphate / Q000378:metabolism; D007295:Inositol Phosphates / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism*; D008870:Microtubules / Q000502:physiology; D054799:Phospholipase C beta; D011976:Receptors, Muscarinic / Q000502:physiology*; D011994:Recombinant Proteins; D015398:Signal Transduction; D018411:Spodoptera; D014404:Tubulin / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism*
D007295:Inositol Phosphates; D007527:Isoenzymes; D011976:Receptors, Muscarinic; D011994:Recombinant Proteins; D014404:Tubulin; D006165:Guanylyl Imidodiphosphate; C091533:glycerophosphoinositol 4,5-bisphosphate; D010738:Type C Phospholipases; D054799:Phospholipase C beta; D019204:GTP-Binding Proteins