Abstract
The Arabidopsis proline-rich, extensin-like receptor-like kinases (PERKs) are a small group of receptor-like kinases that are thought to act as sensors at the cell wall through their predicted proline-rich extracellular domains. In this study, we focused on the characterization of a subclade of three Arabidopsis predicted PERK genes, PERK8, -9, and -10, for which no functions were known. Yeast two-hybrid interaction studies were conducted with the PERK8,- 9, and -10 cytosolic kinase domains, and two members of the Arabidopsis AGC VIII kinase family were identified as interacting proteins: AGC1-9 and the closely related kinesin-like calmodulin-binding protein (KCBP)-interacting protein kinase (KIPK). As KIPK has been identified previously as an interactor of KCBP, these interactions were also examined further and confirmed in this study. Finally, T-DNA mutants for each gene were screened for altered phenotypes under different conditions, and from these screens, a role for the PERK, KIPK, and KCBP genes in negatively regulating root growth was uncovered.
Highlights
Plant receptor-like kinases make up the largest class of kinases in plant genomes, with over 600 predicted members in Arabidopsis, and have a large range of functions from plant development to plant–microbe interactions and abiotic stress responses
Yeast two-hybrid interaction studies were conducted with the PERK8, 9, and -10 cytosolic kinase domains, and two members of the Arabidopsis AGC VIII kinase family were identified as interacting proteins: AGC1-9 and the closely related kinesin-like calmodulin-binding protein (KCBP)-interacting protein kinase (KIPK)
These constructs were first tested for background activity, and from this, PERK10 was chosen for the full-scale screen of a yeast two-hybrid library constructed of random-primed cDNA from Arabidopsis flower bud mRNA
Summary
Plant receptor-like kinases make up the largest class of kinases in plant genomes, with over 600 predicted members in Arabidopsis, and have a large range of functions from plant development to plant–microbe interactions and abiotic stress responses (reviewed by Gish and Clark, 2011; Antolin-Llovera et al, 2012; Osakabe et al, 2013). One predicted subclass of the plant receptor-like kinases is the proline-rich, extensinlike receptor-like kinases (PERKs) (Silva and Goring, 2002) With their predicted proline-rich, extensin-like extracellular domains, PERKs are thought to be part of a group of proteins that act as sensors/receptors at the cell wall. These sensors/receptors, for example, may monitor changes to the cell wall during cell expansion in plant growth, or during plant exposure to abiotic/biotic stresses, and activate associated cellular responses (reviewed by Humphrey et al, 2007; Steinwand and Kieber, 2010; Doblin et al, 2014). Arabidopsis PERK4 has been shown to have kinase activity, and a GFP:PERK4 fusion was localized to the plasma membrane and appeared to be associated with the cell wall (Bai et al, 2009)
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