Abstract
The plastid potassium cation efflux antiporters (KEAs) are important for chloroplast function, development, and photosynthesis. To understand their regulation at the protein level is therefore of fundamental importance. Prior studies have focused on the regulatory K+ transport and NAD-binding (KTN) domain in the C-terminus of the thylakoid carrier KEA3 but the localization of this domain remains unclear. While all three plastid KEA members are highly conserved in their transmembrane region and the C-terminal KTN domain, only the inner envelope KEA family members KEA1 and KEA2 carry a long soluble N-terminus. Interestingly, this region is acetylated at lysine 168 by the stromal acetyltransferase enzyme NSI. If an odd number of transmembrane domains existed for inner envelope KEAs, as it was suggested for all three plastid KEA carriers, regulatory domains and consequently protein regulation would occur on opposing sides of the inner envelope. In this study we therefore set out to investigate the topology of inner envelope KEA proteins. Using a newly designed antibody specific to the envelope KEA1 N-terminus and transgenic Arabidopsis plants expressing a C-terminal KEA1–YFP fusion protein, we show that both, the N-terminal and C-terminal, regulatory domains of KEA1 reside in the chloroplast stroma and not in the intermembrane space. Considering the high homology between KEA1 and KEA2, we therefore reason that envelope KEAs must consist of an even number of transmembrane domains.
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Acknowledgements
Funding: BB DFG, CRC TR175 (B06), SS DFG, CRC 1035 (A04), H-HK NSF Career Award (IOS-1553506), 3rd call ERA-CAPS funding from NSF (IOS-1847382), and (#DE-SC0017160) from DOE. We thank Professor Dr. Jürgen Soll from LMU for supporting this project by initiating the production of α-KEA1(2).
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Bölter, B., Mitterreiter, M.J., Schwenkert, S. et al. The topology of plastid inner envelope potassium cation efflux antiporter KEA1 provides new insights into its regulatory features. Photosynth Res 145, 43–54 (2020). https://doi.org/10.1007/s11120-019-00700-2
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DOI: https://doi.org/10.1007/s11120-019-00700-2