Elevated Rac1 activity changes the M3 muscarinic acetylcholine receptor-mediated inhibition of proliferation to induction of cell death.
Although muscarinic acetylcholine receptors (mAChRs) regulate proliferation in many cell types, the signaling pathways involved are unclear. The participation of the small GTPases Rac1 and RhoA in M(3) mAChR-mediated inhibition of proliferation was investigated by activating M(3) mAChRs stably transfected in Chinese hamster ovary cells stably coexpressing hemagglutinin (HA)-tagged wild-type or mutant Rac1 or RhoA proteins. Activation of M(3) mAChRs activates both Rac1 and RhoA and inhibits cell proliferation in all cell lines tested. mAChR-mediated inhibition of proliferation is diminished in cells expressing dominant-negative HA-Rac1(Asn17) (m3DNRac) but is enhanced in cells expressing HA-Rac1 (m3WTRac) or constitutively active HA-Rac(Val12) (m3CARac). The activation of mAChRs in m3WTRac and m3CARac cells also induces apoptosis. Expression of wild-type or mutant RhoA proteins does not alter mAChR-mediated inhibition of proliferation. mAChR-induced inhibition of proliferation is abrogated in all cell lines when Galpha(q/11) signaling is terminated by transient expression of the COOH-terminal fragment of phospholipase C (PLC-beta1ct), the NH(2)-terminal fragment of G protein-coupled receptor kinase, or the regulator of G protein signaling 2. Pretreatment of all cells expressing wild-type or mutant Rac1 proteins with edelfosine, a phosphatidylinositol-specific PLC inhibitor, or Go 6976, which inhibits conventional protein kinase C (PKC) isoforms, diminishes the M(3) mAChR's ability to inhibit proliferation. Our results identify Galpha(q/11), PLC, and PKC as participants in the M(3) mAChR-mediated inhibition of cell proliferation. These findings indicate that in the context of high Rac1 activity, but not RhoA activity, M(3) mAChR-mediated activation of these participants triggers cell death.
Shafer
Shulamith H
SH
Molecular Pharmacology Laboratory, One Guthrie Square, Sayre, PA 18840, USA.
Williams
Carol L
CL
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D016466:CHO Cells; D016923:Cell Death / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D006224:Cricetinae; D005260:Female; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism; D015870:Gene Expression; D007365:Intercellular Junctions / Q000502:physiology; D007527:Isoenzymes / Q000502:physiology; D011493:Protein Kinase C / Q000502:physiology; D043643:Receptor, Muscarinic M1 / Q000502:physiology; D043587:Receptor, Muscarinic M3 / Q000502:physiology*; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000502:physiology; D020830:rac1 GTP-Binding Protein / Q000378:metabolism* / Q000502:physiology; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
D007527:Isoenzymes; D043643:Receptor, Muscarinic M1; D043587:Receptor, Muscarinic M3; D011493:Protein Kinase C; D010738:Type C Phospholipases; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020830:rac1 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein