Injection of depolarizing current into vegetative cells of the water mold Blastocladiella emersonii elicits a regenerative response that has the electrical characteristics of an action potential. Once they have been taken past a threshold of about -40 mV, cells abruptly depolarize to +20 mV or above; after an interval ranging from several hundred milliseconds to a few seconds, the cells spontaneously return to their resting potential near -100 mV. When the action potential was analyzed with voltage-clamp recording, it proved to be biphasic. The initial phase reflects an influx of calcium ions through voltage-sensitive channels that also carry Sr2+ ions. The delayed, and more extended, phase of inward current results from the efflux of chloride and other anions. The anion channels are broadly selective, passing chloride, nitrate, phosphate, acetate, succinate and even PIPES. The anion channels open in response to the entry of calcium ions, but do not recognize Sr2+. Calcium channels, anion channels and calcium-specific receptors that link the two channels appear to form an ensemble whose physiological function is not known. Action potentials rarely occur spontaneously but can be elicited by osmotic downshock, suggesting that the ion channels may be involved in the regulation of turgor.

Action Potentials↗

PubMed1982

Circulation of potassium across the plasma membrane of Blastocladiella emersonii: K+ channel.

A previous paper reported that the water mold Blastocladiella emersonii generates a transcellular electrical current, such that positive charges enter the rhizoid and leave from the thallus (Stump et al., Proc. Natl. Acad. Sci. U.S.A. 77: 6673-6677, 1980). To begin to understand the genesis of this current we investigated ionic relationships in this organism by use of intracellular microelectrodes. In cells suspended in buffered CaCl2, the membrane potential could be accounted for as a K+ diffusion potential; no evidence for an electrogenic pump was obtained. Potassium ions diffuse outward by a pathway that also carries Rb+ and Ba2+, but excludes both smaller and larger ions (Li+, Na+, Cs+, Mg2+, Ca2+, and choline). Chloride and other anions make little contribution to the potential, but the presence of Ca2+ in the external medium is required for successful potential measurements. In growing cells, the internal K+ concentration is generally somewhat higher than would be expected if the K+ distribution were determined entirely by the membrane potential. Under certain conditions, net uptake of K+ against the electrochemical potential gradient was observed. We suggest that K+ is actively accumulated by a primary transport system that may exchange K+ for H+, and that K+ leaks passively outward through the K+ channel. The K+ circulation across the membrane amounts to about 2% of the K+ pool per min, or 4.5 microA/cm2 of surface area. We propose that this K+ circulation is one arm of the transcellular current, carrying positive charge out of the thallus.

Anions↗

PubMed1980

Ionic control of germination of Blastocladiella emersonii zoospores.

Encystment and germination of Blastocladiella emersonii zoospores involve a rapid and radical transformation of the motile but nongrowing spore into a sessile, growing germling. Certain inorganic ions, notably 50 mM KCl, are efficient inducers of germination. By use of the carbocyanine dye DiO-C6-(3), we found that KCl depolarizes the plasma membrane of zoospores and noted good correlation between depolarization and subsequent germination. Zoospores avidly accumulated K+ ions from the medium, attaining an internal concentration of over 50 mM and a concentration gradient of 2,500. Sodium ions, by contrast, were expelled. Internal K+ was required for normal germination but its function is not known. Zoospores also took up considerable amounts of calcium; most of this was associated with the external surface and appeared to be necessary for maintenance of zoospore integrity. KCl (50 mM) and other salts displaced surface calcium but this was not in itself sufficient to induce germination. The calcium ionophore A23187, in the presence of external calcium, was an effective inducer of germination, suggesting a possible role for cytosolic calcium in triggering the transformation. We propose that the first step in the induction of germination by salts is depolarization of the plasma membrane; subsequent events require the intervention of cytoplasmic signals.

Blastocladiella↗

PubMed1978

ATP-linked calcium transport in cells and membrane vesicles of Streptococcus faecalis.

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Adenosine Triphosphate↗

PubMed1977

Circulation of H+ and K+ across the plasma membrane is not obligatory for bacterial growth.

Streptococcus faecalis grows normally in the presence of gramicidin and other ionophores under conditions such that there is no gradient of pH or of electrical potential across the plasma membrane and that currents of H+, K+, and Na+ are short-circuited. Growth requires a rich medium, a slightly alkaline pH, and a high concentration of external K+. The proton circulation maintains the cytoplasmic pH and pools of ions and other metabolites but is not obligatory for biosynthetic functions including DNA replication, cell division, or assembly of the structural framework of the cell.

Adenosine Triphosphate↗

PubMed1977

Re-examination of F plasmid replication in a dnaC mutant of Escherichia coli.

The replication of an F' plasmid in a dnaC mutant, thermolabile for initiation of chromosomal replication, has been re-examined using a novel DNA-DNA annealing assay. Plasmid replication ceases rapidly at non-permissive conditions, consistent with a direct role for the dnaC product in the replication of F.

Conjugation, Genetic↗

PubMed1968

Identification of a precursor pool of ribosome protein in Escherichia coli.

Antibodies prepared against proteins from 50S ribosomes of Escherichia coli also reacted with the supernatant proteins of a cell-free extract of E. coli which was ribosome-free. A reaction of immunological identity (Ouchterlony tests) was demonstrated for one of these supernatant proteins and one protein found in 50S ribosomes. Isotope experiments involving a shift from (14)C-leucine medium to (12)C-leucine medium showed that these proteins are not formed by breakdown of ribosomes during the preparation of cell-free extracts, but instead represent a pool of ribosome protein which is utilized during growth. In shift experiments from (14)C-leucine to (12)C-leucine medium, the kinetics of disappearance of labeled supernatant ribosome proteins (as measured by reaction with antibody) indicated that half the pool is depleted in 0.1 generation time at 37 C in glucose-salts medium. The pool was also depleted under conditions of amino acid starvation of a "relaxed" strain which accumulated "relaxed" particles. Most, if not all, of the protein present in "relaxed" particles was derived from the pool. The pool represented about 3 to 4% of the total soluble proteins in the ribosome-free supernatant fluid of an E. coli extract.

Bacterial Proteins↗