title: Functional characterization and determination of the physiological role of a calcium-dependent potassium channel from cyanobacteria.
abstract: Despite the important achievement of the high-resolution structures of several prokaryotic channels, current understanding of their physiological roles in bacteria themselves is still far from complete. We have identified a putative two transmembrane domain-containing channel, SynCaK, in the genome of the freshwater cyanobacterium Synechocystis sp. PCC 6803, a model photosynthetic organism. SynCaK displays significant sequence homology to MthK, a calcium-dependent potassium channel isolated from Methanobacterium thermoautotrophicum. Expression of SynCaK in fusion with enhanced GFP in mammalian Chinese hamster ovary cells' plasma membrane gave rise to a calcium-activated, potassium-selective activity in patch clamp experiments. In cyanobacteria, Western blotting of isolated membrane fractions located SynCaK mainly to the plasma membrane. To understand its physiological function, a SynCaK-deficient mutant of Synechocystis sp. PCC 6803, ΔSynCaK, has been obtained. Although the potassium content in the mutant organisms was comparable to that observed in the wild type, ΔSynCaK was characterized by a depolarized resting membrane potential, as determined by a potential-sensitive fluorescent probe. Growth of the mutant under various conditions revealed that lack of SynCaK does not impair growth under osmotic or salt stress and that SynCaK is not involved in the regulation of photosynthesis. Instead, its lack conferred an increased resistance to the heavy metal zinc, an environmental pollutant. A similar result was obtained using barium, a general potassium channel inhibitor that also caused depolarization. Our findings thus indicate that SynCaK is a functional channel and identify the physiological consequences of its deletion in cyanobacteria.
authors: Department of Biology, University of Padua, 35121 Padua, Italy.
authors: Vanessa
authors: V
authors: Checchetto
authors: Elide
authors: E
authors: Formentin
authors: Luca
authors: L
authors: Carraretto
authors: Anna
authors: A
authors: Segalla
authors: Giorgio Mario
authors: GM
authors: Giacometti
authors: Ildiko
authors: I
authors: Szabo
authors: Elisabetta
authors: E
authors: Bergantino
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D005786:Gene Expression Regulation; D008564:Membrane Potentials; D017014:Methanobacterium / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D009997:Osmotic Pressure; D018408:Patch-Clamp Techniques; D024681:Potassium Channels, Calcium-Activated / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D046939:Synechocystis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D024681:Potassium Channels, Calcium-Activated; D011993:Recombinant Fusion Proteins; D015032:Zinc; D002118:Calcium
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