Isolation of a novel G-protein gamma-subunit from Arabidopsis thaliana andits interaction with G beta

There is increasing evidence that heterotrimeric G-proteins (G-proteins) ar e involved in many plant processes including phytohormone response, pathoge n defence and stomatal control. In animal systems, each of the three G-prot ein subunits belong to large multigene families; however, few subunits have been isolated from plants. Here we report the cloning of a second plant G- protein gamma -subunit (AGG2) from Arabidopsis thaliana. The predicted AGG2 protein sequence shows 48% identity to the first identified Arabidopsis G gamma -subunit, AGG1. Furthermore, AGG2 contains all of the conserved chara cteristics of gamma -subunits including a small size (100 amino acids, 11.1 kDa), C-terminal CAAX box and a N-terminal alpha -helix region capable of forming a coiled-coil interaction with the beta -subunit. A strong interact ion between AGG2 and both the tobacco (TGB1) and Arabidopsis (AGB1) beta -s ubunits was observed in vivo using the yeast two-hybrid system. The strong association between AGG2 and AGB1 was confirmed in vitro. Southern and Nort hern analyses showed that AGG2 is a single copy gene in Arabidopsis produci ng two transcripts that are present in all tissues tested. The isolation of a second gamma -subunit from A. thaliana indicates that plant G-proteins, like their mammalian counterparts, may form different heterotrimer combinat ions that presumably regulate multiple signal transduction pathways. (C) 20 01 Elsevier Science B.V. All rights reserved.