A Unique Voltage Sensor Sensitizes the Potassium Channel AKT2 to Phosphoregulation

Among all voltage-gated K+ channels from the model plant Arabidopsis thaliana, the weakly rectifying K+ channel (Kweak channel) AKT2 displays unique gating properties. AKT2 is exceptionally regulated by phosphorylation: when nonphosphorylated AKT2 behaves as an inward-rectifying potassium channel; phosphorylation of AKT2 abolishes inward rectification by shifting its activation threshold far positive (>200 mV) so that it closes only at voltages positive of +100 mV. In its phosphorylated form, AKT2 is thus locked in the open state in the entire physiological voltage range. To understand the molecular grounds of this unique gating behavior, we generated chimeras between AKT2 and the conventional inward-rectifying channel KAT1. The transfer of the pore from KAT1 to AKT2 altered the permeation properties of the channel. However, the gating properties were unaffected, suggesting that the pore region of AKT2 is not responsible for the unique Kweak gating. Instead, a lysine residue in S4, highly conserved among all Kweak channels but absent from other plant K+ channels, was pinpointed in a site-directed mutagenesis approach. Substitution of the lysine by serine or aspartate abolished the “open-lock” characteristic and converted AKT2 into an inward-rectifying channel. Interestingly, phosphoregulation of the mutant AKT2-K197S appeared to be similar to that of the Kin channel KAT1: as suggested by mimicking the phosphorylated and dephosphorylated states, phosphorylation induced a shift of the activation threshold of AKT2-K197S by about +50 mV. We conclude that the lysine residue K197 sensitizes AKT2 to phosphoregulation. The phosphorylation-induced reduction of the activation energy in AKT2 is ∼6 kT larger than in the K197S mutant. It is discussed that this hypersensitive response of AKT2 to phosphorylation equips a cell with the versatility to establish a potassium gradient and to make efficient use of it

HKT2;2/1, a K+-permeable transporter identified in a salt tolerant rice cultivar through surveys of natural genetic polymorphism

We have investigated OsHKT2;1 natural variation in a collection of 49 cultivars with different levels of salt tolerance and geographical origins. The effect of identified polymorphism on OsHKT2;1 activity was analysed through heterologous expression of variants in Xenopus oocytes. OsHKT2;1 appeared to be a highly conserved protein with only five possible amino acid substitutions that have no substantial effect on functional properties. Our study, however, also identified a new HKT isoform, No-OsHKT2;2/1 in Nona Bokra, a highly salt-tolerant cultivar. No-OsHKT2;2/1 probably originated from a deletion in chromosome 6, producing a chimeric gene. Its 5¢ region corresponds to that of OsHKT2;2, whose full-length sequence is not present in Nipponbare but has been identified in Pokkali, a salt-tolerant rice cultivar. Its 3¢ region corresponds to that of OsHKT2;1. No-OsHKT2;2/1 is essentially expressed in roots and displays a significant level of expression at high Na+ concentrations, in contrast to OsHKT2;1. Expressed in Xenopus oocytes or in Saccharomyces cerevisiae, No-OsHKT2;2/1 exhibited a strong permeability to Na+ and K+, even at high external Na+ concentrations, like OsHKT2;2, and in contrast to OsHKT2;1. Our results suggest that No-OsHKT2;2/1 can contribute to Nona Bokra salt tolerance by enabling root K+ uptake under saline conditions

The roots of future rice harvests

The authors thank the Global Rice Science Partnership and Agropolis Fondation (Special grant n° 1400–009 and Rhizopolis grant n° 1001–005) benefiting from a national ANR Investissement d’Avenir” grant ANR-10-LABX-001-01) for supporting the workshop. They acknowledge the assistance of Nathalie Pivot, Cirad and Véronique Rafin, INRA in workshop organization. The root research at Cirad and University of Aberdeen is supported by the European Grant (FP7/2007-2013) under grant agreement n° 289300.27 EURoot “Enhancing resource Uptake from ROOTs under stress in cereal crops”. Research at IRRI is supported by the Generation Challenge Program and the Bill and Melinda Gates Foundation. J.X. is supported by the AcRF Tier 2 grant (MOE2009-T2-1-060) from the Ministry of Education of Singapore and National Research Foundation Singapore under its Competitive Research Programme (CRP Award No. NRF2010 NRF-CRP002-018). Doan Trung Luu is supported by the EU Marie Curie International Outgoing Fellowship 'ORYZAQUA – Cell Biology of Rice Aquaporins' (PIOF-GA-2011-300150). AP acknowledges the Generation Challenge Programme funded project “Targeting drought avoidance root traits to enhance rice productivity under water limited environments”. Financial support for A.G. Diedhiou was provided by the Université Cheikh Anta Diop (UCAD, VE12/13, CpVIII-Ar4 ) and GRISP. *This paper is dedicated to the late memory of Pr Ping Wu who passed away in a tragic car accident on June 12th, 2014.Peer reviewedPublisher PD

CMS physics technical design report : Addendum on high density QCD with heavy ions

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