An intact helical domain is required for Gα14 to stimulate phospholipase Cβ

Abstract

Stimulation of phospholipase Cβ (PLCβ) by the activated α-subunit of G_q (Gα_q) constitutes a major signaling pathway for cellular regulation, and structural studies have recently revealed the molecular interactions between PLCβ and Gα_q. Yet, most of the PLCβ-interacting residues identified on Gα_q are not unique to members of the Gα_q family. Molecular modeling predicts that the core PLCβ-interacting residues located on the switch regions of Gα_q are similarly positioned in Gα_z which does not stimulate PLCβ. Using wild-type and constitutively active chimeras constructed between Gα_z and Gα₁₄, a member of the Gα_q family, we examined if the PLCβ-interacting residues identified in Gα_q are indeed essential.

Four chimeras with the core PLCβ-interacting residues composed of Gα_z sequences were capable of binding PLCβ2 and stimulating the formation of inositol trisphosphate. Surprisingly, all chimeras with a Gα_z N-terminal half failed to functionally associate with PLCβ2, despite the fact that many of them contained the core PLCβ-interacting residues from Gα₁₄. Further analyses revealed that the non-PLCβ2 interacting chimeras were capable of interacting with other effector molecules such as adenylyl cyclase and tetratricopeptide repeat 1, indicating that they could adopt a GTP-bound active conformation.

Collectively, our study suggests that the previously identified PLCβ-interacting residues are insufficient to ensure productive interaction of Gα₁₄ with PLCβ, while an intact N-terminal half of Gα₁₄ is apparently required for PLCβ interaction.