[47] Patch clamp studies of microbial ion channels

Saimi, Yoshiro; Martinac, Boris; Delcour, Anne H.; Minorsky, Peter V.; Gustin, Michael C.; Culbertson, Michael R.; Adler, Julius; Kung, Ching

doi:10.1016/0076-6879(92)07049-t

Cited by 29 publications

(15 citation statements)

References 16 publications

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“…The released vacuoles can then be examined by using a planar bilayer or a patch-clamp pipet. An Ϸ400-pS cation conductance has repeatedly been observed (27,28,31) that is activated by Ca 2ϩ presented from the cytoplasmic side. We cloned the TRP homolog YVC1 of yeast and showed that it is required for this conductance (26).…”

Section: Yvc1p Is Msmentioning

confidence: 99%

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The transient receptor potential channel on the yeast vacuole is mechanosensitive

Zhou

Batiza²,

Loukin³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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Ca 2؉ is released from the vacuole into the yeast cytoplasm on an osmotic upshock, but how this upshock is perceived was unknown. We found the vacuolar channel, Yvc1p, to be mechanosensitive, showing that the Ca 2؉ conduit is also the sensing molecule. Although fragile, the yeast vacuole allows limited direct mechanical examination. Pressures at tens of millimeters of Hg (1 mmHg ‫؍‬ 133 Pa) activate the 400-pS Yvc1p conductance in whole-vacuole recording mode as well as in the excised cytoplasmic-side-out mode. Raising the bath osmolarity activates this channel and causes vacuolar shrinkage and deformation. It appears that, on upshock, a transient osmotic force activates Yvc1p to release Ca 2؉ from the vacuole. Mechanical activation of Yvc1p occurs regardless of Ca 2؉ concentration and is apparently independent of its known Ca 2؉ activation, which we now propose to be an amplification mechanism (Ca 2؉ -induced Ca 2؉ release). Yvc1p is a member of the transient receptor potential-family channels, several of which have been associated with mechanosensation in animals. The possible use of Yvc1p as a molecular model to study mechanosensation in general is discussed.

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Section: Yvc1p Is Msmentioning

confidence: 99%

“…Vacuoles were generated as described (27,28). The procedure entails an overnight culturing of the spheroplasts, in which the multivesicular looking vacuole(s) in each logarithmic-phase cell (ref.…”

Section: Methodsmentioning

confidence: 99%

The transient receptor potential channel on the yeast vacuole is mechanosensitive

Zhou

Batiza²,

Loukin³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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145

123

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“…These techniques are similar to those we have described [6,7,17]. Briefly, cells were collected by centrifugation and the cell pellet was resuspended and incubated at 30°C for 20 min with 0.13 mg/ml Zymolyase (ICN) in 0.4 M sorbitol to partially digest the cell wall while supporting the emerging spheroplasts osmotically.…”

Section: Patch Clamp Techniquesmentioning

confidence: 99%

YKC1 encodes the depolarization‐activated K⁺ channel in the plasma membrane of yeast

et al. 1995

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“…The Yvc1 channel was first identified by electrophysiological studies of ion conductances in isolated yeast vacuoles (Bertl and Slayman 1990;Saimi et al 1992), which showed that it was cation selective and inhibited by vacuolar Ca 2+ (mM) or low pH (,5). Many years later, the gene product responsible for this conductance was identified as the product of the Yvc1 gene, which is a member of the large family of TRP channels (transient receptor potential), whose mammalian members are responsible for sensory phenomenon such as detection of heat, cold, and pain and are associated with diseases including retinal degeneration and polycystic kidney disease (Palmer et al 2001;Nilius and Owsianik 2011).…”

Section: Ca 2+ Release From Intracellular Compartmentsmentioning

confidence: 99%

Regulation of Cation Balance inSaccharomyces cerevisiae

2013

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All living organisms require nutrient minerals for growth and have developed mechanisms to acquire, utilize, and store nutrient minerals effectively. In the aqueous cellular environment, these elements exist as charged ions that, together with protons and hydroxide ions, facilitate biochemical reactions and establish the electrochemical gradients across membranes that drive cellular processes such as transport and ATP synthesis. Metal ions serve as essential enzyme cofactors and perform both structural and signaling roles within cells. However, because these ions can also be toxic, cells have developed sophisticated homeostatic mechanisms to regulate their levels and avoid toxicity. Studies in Saccharomyces cerevisiae have characterized many of the gene products and processes responsible for acquiring, utilizing, storing, and regulating levels of these ions. Findings in this model organism have often allowed the corresponding machinery in humans to be identified and have provided insights into diseases that result from defects in ion homeostasis. This review summarizes our current understanding of how cation balance is achieved and modulated in baker's yeast. Control of intracellular pH is discussed, as well as uptake, storage, and efflux mechanisms for the alkali metal cations, Na + and K + , the divalent cations, Ca 2+ and Mg 2+ , and the trace metal ions, Fe 2+ , Zn 2+ , Cu 2+ , and Mn 2+ . Signal transduction pathways that are regulated by pH and Ca 2+ are reviewed, as well as the mechanisms that allow cells to maintain appropriate intracellular cation concentrations when challenged by extreme conditions, i.e., either limited availability or toxic levels in the environment. TABLE OF CONTENTS Abstract 677Introduction 678

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[47] Patch clamp studies of microbial ion channels

Cited by 29 publications

References 16 publications

The transient receptor potential channel on the yeast vacuole is mechanosensitive

The transient receptor potential channel on the yeast vacuole is mechanosensitive

YKC1 encodes the depolarization‐activated K⁺ channel in the plasma membrane of yeast

Regulation of Cation Balance inSaccharomyces cerevisiae

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[47] Patch clamp studies of microbial ion channels

Cited by 29 publications

References 16 publications

The transient receptor potential channel on the yeast vacuole is mechanosensitive

The transient receptor potential channel on the yeast vacuole is mechanosensitive

YKC1 encodes the depolarization‐activated K+ channel in the plasma membrane of yeast

Regulation of Cation Balance inSaccharomyces cerevisiae

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Product

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YKC1 encodes the depolarization‐activated K⁺ channel in the plasma membrane of yeast