Signaling cascades mediated by heterotrimeric G proteins are ubiquitous and important signal transduction mechanisms in both metazoans and plants. In the model plant Arabidopsis thaliana, the sole canonical G protein α subunit, GPA1, has been implicated in multiple signaling events, including guard cell movement regulated by the plant stress hormone abscisic acid (ABA). However, only a handful of proteins have been demonstrated to be involved in GPA1 signaling to date. Here, we compared the proteome composition of guard cells from wild type Col vs gpa1-4 null mutants with and without ABA treatment using iTRAQ technology to identify guard cell proteins whose abundance was affected by ABA and/or GPA1. After imposition of strict selection criteria, the abundance of two proteins in Col and six proteins in gpa1-4 was found to be affected by ABA in guard cells, and 18 guard cell proteins were quantitatively affected by the mutation of GPA1. On the basis of known functions of the differentially expressed proteins, our data suggest that GPA1 inhibits guard cell photosynthesis and promotes the availability of reactive oxygen species (ROS) in guard cells. These results exemplify how iTRAQ can be used to quantitatively study single cell signaling pathways in Arabidopsis.
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