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Proc Natl Acad Sci U S A
2009 Dec 15;10650:21425-30. doi: 10.1073/pnas.0912021106.
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Activity of guard cell anion channel SLAC1 is controlled by drought-stress signaling kinase-phosphatase pair.
Geiger D, Scherzer S, Mumm P, Stange A, Marten I, Bauer H, Ache P, Matschi S, Liese A, Al-Rasheid KA, Romeis T, Hedrich R.
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In response to drought stress the phytohormone ABA (abscisic acid) induces stomatal closure and, therein, activates guard cell anion channels in a calcium-dependent as well as-independent manner. Two key components of the ABA signaling pathway are the protein kinase OST1 (open stomata 1) and the protein phosphatase ABI1 (ABA insensitive 1). The recently identified guard cell anion channel SLAC1 appeared to be the key ion channel in this signaling pathway but remained electrically silent when expressed heterologously. Using split YFP assays, we identified OST1 as an interaction partner of SLAC1 and ABI1. Upon coexpression of SLAC1 with OST1 in Xenopus oocytes, SLAC1-related anion currents appeared similar to those observed in guard cells. Integration of ABI1 into the SLAC1/OST1 complex, however, prevented SLAC1 activation. Our studies demonstrate that SLAC1 represents the slow, deactivating, weak voltage-dependent anion channel of guard cells controlled by phosphorylation/dephosphorylation.
Fig. 2. Interactions between proteins involved in the ABA signaling pathway by bimolecular fluorescence complementation (BIFC) in Xenopus oocytes (A–D) as well as Arabidopsis mesophyll protoplasts (E) and (F). Pictures (A–D), taken with a confocal laser scanning microscope, show a quarter of an optical slice of an oocyte (see inset in (C). (A) Noninjected control oocyte. (B) SLAC1::YFPC coexpressed with YFPN. (C) SLAC1::YFPC coexpressed with OST1::YFPN. (D) ABI1::YFPC coexpressed with OST1::YFPN. (E and F) Interaction of SLAC1 and OST1, as well as ABI1 and OST1, monitored in transiently transformed Arabidopsis mesophyll protoplasts by bimolecular fluorescence complementation. (Left) Overlay of transmitted light and chlorophyll fluorescence. (Right) Overlay of YFP and chlorophyll fluorescence. Plasmid combinations were as follows: (E) YFPC::SLAC1 plus YFPN::OST1 and (F) YFPC::ABI1 plus YFPN::OST1.
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