The <i>Saccharomyces cerevisiae</i> CCH1 gene is involved in calcium influx and mating

Fischer, Marc; Schnell, Norbert; Chattaway, J. A.; Davies, Paul; Dixon, Graham K.; Sanders, Dale

doi:10.1016/s0014-5793(97)01466-x

Cited by 166 publications

(180 citation statements)

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“…These non-pore-forming subunits dramatically influence the properties and surface expression of the channels (12)(13)(14)(15)(16). Although the channel activity of Cch1 has not yet been revealed experimentally and Mid1 has no homology to the auxiliary subunits, Mid1 has been shown to cooperate with Cch1 in mating pheromone-induced Ca 2ϩ uptake (8,9,17), store-operated or capacitative Ca 2ϩ entry (10), endoplasmic reticulum stress-induced Ca 2ϩ uptake (18), and a hyperosmotic stress-induced increase in cytosolic Ca 2ϩ (19). On the other hand, it has been shown recently that Mid1, but not Cch1, is required for an antiarrhythmic drug amiodarone-induced increase in cytosolic Ca 2ϩ mainly caused by Ca 2ϩ influx (20).…”

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confidence: 99%

Molecular Dissection of the Hydrophobic Segments H3 and H4 of the Yeast Ca2+ Channel Component Mid1

Tada¹,

Ohmori²,

Iida³

2003

Journal of Biological Chemistry

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The Saccharomyces cerevisiae MID1 gene product, Mid1, is composed of 548 amino acid residues, has four relatively hydrophobic segments named H1-H4, and functions as a Ca 2؉ -permeable, stretch-activated channel when expressed in mammalian cells. In some conditions Mid1 cooperates with Cch1, a yeast homolog of the ␣1 subunit of mammalian voltage-gated channels. To identify the important regions or amino acid residues necessary for Mid1 function, we employed in vitro sitedirected mutagenesis on H3 and H4 of Mid1 and expressed the resulting mutant genes in a mid1 null mutant to examine whether the mutant gene products are functional or not in vivo. Mutant Mid1 proteins lacking the whole H3 or H4 segment, H3De or H4De, did not complement the lethality and low Ca 2؉ accumulation activity of the mid1 mutant, although their localization and contents appeared to be normal, indicating that H3 and H4 are required for Mid1 function itself. Single amino acid exchange experiments on individual amino acid residues of H3 and H4 showed that 10 of 20 residues in H3 and 14 of 23 residues in H4 were important for the normal function of Mid1. In particular, we found four severe loss-of-function mutations, D341E, F356S, C373D, and C373R, and two interesting mutations leading to a high level of Ca 2؉ accumulation with a slightly low complementing activity, G342A and Y355A. The importance of these amino acid residues will be discussed.

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confidence: 99%

Molecular Dissection of the Hydrophobic Segments H3 and H4 of the Yeast Ca2+ Channel Component Mid1

Tada¹,

Ohmori²,

Iida³

2003

Journal of Biological Chemistry

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“…In G. zeae, the double mutant of both channel proteins exhibited a stronger phenotype than the single mid1 deletion mutant (26), supporting the idea of both dependent and independent roles for Mid1 and Cch1. In B. cinerea, the single and double knockout mutants displayed similar phenotypes under all conditions tested, suggesting that these two proteins act in a complex and have common functions, as also reported for mid1 and cch1 knockout mutants in Candida albicans and S. cerevisiae (21)(22)(23)58). To study a putative interaction between these two proteins in B. cinerea, we used a split ubiquitinbased yeast two-hybrid system (Dualsystems).…”

Section: Discussionmentioning

confidence: 58%

“…In yeast, Cch1 is localized in the plasma membrane, where it forms a complex with Mid1. Both proteins have been shown to be required for the uptake of extracellular Ca 2ϩ in cells responding to mating pheromones (13,(21)(22)(23)(24).…”

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Cch1 and Mid1 Are Functionally Required for Vegetative Growth under Low-Calcium Conditions in the Phytopathogenic Ascomycete Botrytis cinerea

Harren

Tudzynski

2013

Eukaryot Cell

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In the filamentous phytopathogen Botrytis cinerea, the Ca 2؉ /calcineurin signaling cascade has been shown to play an important role in fungal growth, differentiation, and virulence. This study deals with the functional characterization of two components of this pathway, the putative calcium channel proteins Cch1 and Mid1. The cch1 and mid1 genes were deleted, and single and double knockout mutants were analyzed during different stages of the fungal life cycle. Our data indicate that Cch1 and Mid1 are functionally required for vegetative growth under conditions of low extracellular calcium, since the growth of both deletion mutants is strongly impaired when they are exposed to the Ca 2؉ -chelating agents EGTA and 1,2-bis(o-aminophenoxy)ethane-N,N,N=,N=-tetraacetic acid (BAPTA). The impact of external Ca 2؉ was investigated by supplementing with CaCl 2 and the ionophore A23187, both of which resulted in elevated growth for all mutants. However, deletion of either gene had no impact on germination, sporulation, hyphal morphology, or virulence. By use of the aequorin reporter system to measure intracellular calcium levels, no differences between the mutant strains and the wild type were obtained. Localization studies revealed a subcellular distribution of the Mid1-green fluorescent protein (GFP) fusion protein in network-like filaments, probably the endoplasmic reticulum (ER) membranes, indicating that Mid1 is not a plasma membrane-located calcium channel in B. cinerea.

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“…albicans (Brand et al, 2007) and Sac. cerevisiae (Fischer et al, 1997;Iida et al, 2004;Paidhungat & Garrett, 1997).…”

Section: Discussionmentioning

confidence: 99%

Deletion of Mid1, a putative stretch-activated calcium channel in Claviceps purpurea, affects vegetative growth, cell wall synthesis and virulence

Bormann

Tudzynski

2009

Microbiology

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Deletion of Mid1, a putative stretch-activated calcium channel in Claviceps purpurea, affects vegetative growth, cell wall synthesis and virulence The putative Claviceps purpurea homologue of the Saccharomyces cerevisiae stretch-activated calcium ion channel Mid1 was investigated for its role in vegetative growth, differentiation and pathogenicity on rye (Secale cereale). Gene replacement mutants of Cl. purpurea mid1 were not affected in polar growth and branching in axenic culture but showed a significantly reduced growth rate. The growth defect could not be complemented by Ca 2+ supplementation, in contrast to mid1 mutants in yeast, but the altered sensitivity of the mutants to changes in external and internal Ca 2+ concentrations indicates some role of Mid1 in Ca 2+ homeostasis. The major effect of mid1 deletion, however, was the complete loss of virulence: infected rye plants showed no disease symptoms at all. Detailed analyses of in vitro-infected rye ovaries demonstrated that the Dmid1 mutants had multiple apical branches and were unable to infect the host tissue, suggesting that Mid1 is essential for generating the necessary mechanical force for penetration. This is believed to be the first report of an essential role for a Mid1 homologue in the virulence of a plantpathogenic fungus. INTRODUCTIONThe phytopathogenic biotrophic ascomycete Claviceps purpurea (Fries ex Fries) Tulasne parasitizes a large number of Poaceae species (Tudzynski & Scheffer, 2004). It colonizes ovaries of its main host, rye (Secale cereale), by penetrating the cuticle and growing through the ovarian tissue in a strictly directed and unbranched manner until it reaches the vascular bundles. By tapping into the vascular bundles, the fungus assures a stable nutrition supply. This is a prerequisite for the subsequent colonization of the entire ovary. During this process the fungus produces masses of conidia which are secreted in a viscous fluid called honeydew. Spores are dispersed to other blooming florets mainly zoochorically and hydrochorically. The period of conidia formation ends when sclerotium formation is initiated. The sclerotia represent resting structures consisting of a solid plectenchymatous structure of pigmented storage cells (Tenberge, 1999). They contain the so-called ergot alkaloids, which represent a class of pharmacologically important mycotoxins (Schardl et al., 2006).The fungus grows mainly intercellularly and therefore decomposes the middle lamella between the plant cells. This lamella predominantly consists of pectin; degradation of pectin by fungal polygalacturonases is essential for full pathogenicity (Oeser et al., 2002). It is possible that Ca 2+ complexed in this pectin could act as a trail for the hyphae on their way through the ovary. A tip-high gradient of cytoplasmic calcium ([Ca 2+ ] cyt. ) has been observed in a wide range of eukaryotes that grow in a polarized and oriented manner, including fungal hyphae Silverman-Gavrila & Lew, 2003), neurons (Mattson, 1999), and pollen tubes and root hairs of vascula...

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The Saccharomyces cerevisiae CCH1 gene is involved in calcium influx and mating

Cited by 166 publications

References 16 publications

Molecular Dissection of the Hydrophobic Segments H3 and H4 of the Yeast Ca2+ Channel Component Mid1

Molecular Dissection of the Hydrophobic Segments H3 and H4 of the Yeast Ca2+ Channel Component Mid1

Cch1 and Mid1 Are Functionally Required for Vegetative Growth under Low-Calcium Conditions in the Phytopathogenic Ascomycete Botrytis cinerea

Deletion of Mid1, a putative stretch-activated calcium channel in Claviceps purpurea, affects vegetative growth, cell wall synthesis and virulence

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