Woychik NA et al. (1990) Genes Dev "Subunits shared by eukaryotic nuclear RNA polymerases."

Liao SM, Woychik NA, Kolodziej PA, Young RA

[Genes Dev, 1990]

RNA polymerases I, II, and III share three subunits that are immunologically and biochemically indistinguishable. The Saccharomyces cerevisiae genes that encode these subunits (RPB5, RPB6, and RPB8) were isolated and sequenced, and their transcriptional start sites were deduced. RPB5 encodes a 25-kD protein, RPB6, an 18-kD protein, and RPB8, a 16-kD protein. These genes are single copy, reside on different chromosomes, and are essential for viability. The fact that the genes are single copy, corroborates previous evidence suggesting that each of the common subunits is identical in RNA polymerases I, II, and III. Furthermore, immunoprecipitation of RPB6 coprecipitates proteins whose sizes are consistent with RNA polymerase I, II, and III subunits. Sequence similarity between the yeast RPB5 protein and a previously characterized human RNA polymerase subunit demonstrates that the common subunits of the nuclear RNA polymerases are well conserved among eukaryotes. The presence of these conserved and essential subunits in all three nuclear RNA polymerases and the absence of recognizable sequence motifs for DNA and nucleoside triphosphate-binding indicate that the common subunits do not have a catalytic role but are important for a function shared by the RNA polymerases such as transcriptional efficiency, nuclear localization, enzyme stability, or coordinate regulation of rRNA, mRNA, and tRNA synthesis.

Zou J et al. (2023) Nat Biotechnol "A DNA nanodevice for mapping sodium at single-organelle resolution."

Gupta PD, Mitra K, Smith JJ, Rao R, Ramirez JR, Zou J, Anees P, Krishnan Y, Oettinger D, Veetil AT, Kratsios P

[Nat Biotechnol, 2023]

Cellular sodium ion (Na⁺) homeostasis is integral to organism physiology. Our current understanding of Na⁺ homeostasis is largely limited to Na⁺ transport at the plasma membrane. Organelles may also contribute to Na⁺ homeostasis; however, the direction of Na⁺ flow across organelle membranes is unknown because organellar Na⁺ cannot be imaged. Here we report a pH-independent, organelle-targetable, ratiometric probe that reports lumenal Na⁺. It is a DNA nanodevice containing a Na⁺-sensitive fluorophore, a reference dye and an organelle-targeting domain. By measuring Na⁺ at single endosome resolution in mammalian cells and Caenorhabditis elegans, we discovered that lumenal Na⁺ levels in each stage of the endolysosomal pathway exceed cytosolic levels and decrease as endosomes mature. Further, we find that lysosomal Na⁺ levels in nematodes are modulated by the Na⁺/H⁺ exchanger NHX-5 in response to salt stress. The ability to image subcellular Na⁺ will unveil mechanisms of Na⁺ homeostasis at an increased level of cellular detail.

Yuan A et al. (2003) Neuron "The sodium-activated potassium channel is encoded by a member of the Slo gene family."

Yuan A, Santi CM, Pollak K, Kaczmarek L, Crowder CM, Wang ZW, Wei A, Nonet M, Salkoff L

[Neuron, 2003]

Na+-activated potassium channels (K-Na) have been identified in cardiomyocytes and neurons where they may provide protection against ischemia. We now report that K-Na is encoded by the rSlo2 gene (also called Slack), the mammalian ortholog of slo-2 in C. elegans. rSlo2, heterologously expressed, shares many properties of native K-Na including activation by intracellular Na+, high conductance, and prominent subconductance states. In addition to activation by Na+, we report that rSLO-2 channels are cooperatively activated by intracellular Cl-, similar to C. elegans SLO-2 channels. Since intracellular Na+ and Cl- both rise in oxygen-deprived cells, coactivation may more effectively trigger the activity of rSLO-2 channels in ischemia. In C. elegans, mutational and physiological analysis revealed that the SLO-2 current is a major component of the delayed rectifier. We demonstrate in C. elegans that slo-2 mutants are hypersensitive to hypoxia, suggesting a conserved role for the slo-2 gene

Wang L et al. (2017) J Neurosci "A Gustatory Neural Circuit of Caenorhabditis elegans Generates Memory-Dependent Behaviors in Na+ Chemotaxis."

Wang L, Sato H, Kunitomo H, Tomioka M, Satoh Y, Iino Y

[J Neurosci, 2017]

Animals show various behaviors in response to environmental chemicals. These behaviors are often plastic depending on previous experiences. Caenorhabditis elegans, which has highly developed chemosensory system with a limited number of sensory neurons, is an ideal model for analyzing the role of each neuron in innate and learned behaviors. Here we report a new type of memory-dependent behavioral plasticity in Na(+) chemotaxis generated by the left member of bilateral gustatory neuron pair ASE (ASEL neuron). When worms were cultivated in the presence of Na(+), they showed positive chemotaxis towards Na(+); but when worms were cultivated under Na(+)-free conditions, they showed no preference in Na(+) concentration. Both channelrhodopsin-2 (ChR2) activation with blue light and upsteps of Na(+) concentration activated ASEL only after cultivation with Na(+), as judged by increase in intracellular Ca(2+) Under cultivation conditions with Na(+), photoactivation of ASEL caused activation of its downstream interneurons AIY and AIA, which stimulate forward locomotion, and inhibition of its downstream interneuron AIB, which inhibits the turning/reversal behavior, and overall drove worms towards higher Na(+) concentrations. We also found that the Gq signaling pathway and the neurotransmitter glutamate are both involved in the behavioral response generated by ASEL. SIGNIFICANCE STATEMENT: Animals have acquired various types of behavioral plasticity during their long evolutionary history. C. elegans prefers odors associated with food, but plastically changes its behavioral response according to previous experience. Here we report a new type of behavioral response generated by a single gustatory sensory neuron, the ASE-left neuron (ASEL). ASEL did not respond to photostimulation or upsteps of Na(+) concentration when worms were cultivated in Na(+)-free conditions; however, when worms were cultivated with Na(+), ASEL responded and inhibited AIB to avoid turning, and stimulated AIY and AIA to promote forward locomotion, which collectively drove worms towards higher Na(+) concentrations. Glutamate and the Gq signaling pathway are essential for driving worms towards higher Na(+) concentrations.

Johnson CK et al. (2020) J Neurophysiol "The Na+/K+-ATPase is needed in glia of touch receptors for responses to touch in C. elegans."

Wang Y, Bianchi L, Fernandez-Abascal J, Johnson CK, Wang L

[J Neurophysiol, 2020]

Four of the five types of mammalian mechanosensors are composed of nerve endings and accessory cells. In C. elegans we showed that glia supports the function of nose touch neurons via the activity of glial Na⁺ and K⁺channels. We show here that a third regulator of Na⁺ and K⁺, the Na⁺/K⁺-ATPase, is needed in glia of nose touch neurons for touch. Importantly, we show that the two Na⁺/K⁺-ATPase genes are needed for the function rather than structural integrity and that their ion transport activity is crucial for touch. Finally, when glial Na⁺/K⁺-ATPase genes are knocked-out, touch can be restored by activation of a third Na⁺/K⁺-ATPase. Taken together, these data show the requirement in glia of touch neurons of the function of the Na⁺/K⁺-ATPase. These data underscore the importance of the homeostasis of Na⁺ and K⁺, most likely in the space surrounding touch neurons, in touch sensation, a function that might be conserved across species.

Bruckbauer A et al. (2017) Am J Cardiovasc Dis "Leucine-nicotinic acid synergy stimulates AMPK/Sirt1 signaling and regulates lipid metabolism and lifespan in Caenorhabditis elegans, and hyperlipidemia and atherosclerosis in mice."

Zemel MB, Li F, Cao Q, Cui X, Bruckbauer A, Zha L, Jing J, Xue B, Banerjee J, Shi H

[Am J Cardiovasc Dis, 2017]

BACKGROUND/AIMS: Nicotinic acid (NA), a lipid-lowering drug, serves as a source of NAD(+), the cofactor for Sirt1. Leucine (Leu) stimulates the AMPK/Sirt1 axis and amplifies the effects of other AMPK/Sirt1 activating compounds. Therefore, we tested the interactive effects of leucine and low dose NA on AMPK/Sirt1 signaling and downstream effects of lipid metabolism in cell culture, C. elegans and mice. METHODS: LDL-receptor knockout mice were fed an atherogenic Western diet supplemented with leucine (24 g/kg diet) and sub-therapeutic NA combinations (50 mg/kg diet and 250 mg/kg diet) or low therapeutic NA (1000 mg/kg diet) for 8 weeks to evaluate markers of hyperlipidemia and atherosclerosis. RESULTS: NA-Leu increased P-AMPK and Sirt1 in adipocytes and myotubes. In C. elegans, NA-Leu increased P-AMPK and DAF-16 (FOXO), reduced lipid accumulation and increased median survival under mild oxidative stress conditions. In the mice, NA-Leu reduced total cholesterol, cholesterol esters, plasma triglycerides, atherosclerotic lesion size, lipid area, and aortic macrophage infiltration, similar to the therapeutic NA dose. CONCLUSION: Leu amplifies the effects of NA on lipid metabolism, hyperlipidemia and atherosclerosis in mice, at least in part by activation of the AMPK/Sirt1 axis. This combination may be a potential therapeutic alternative for hyperlipidemia and atherosclerosis.

Sharma V et al. (2016) Bioinform Biol Insights "Nematode Sodium Calcium Exchangers: A Surprising Lack of Transport."

O'Halloran DM, Sharma V

[Bioinform Biol Insights, 2016]

Na(+)/Ca(2+) exchangers are low-affinity, high-capacity transporters that rapidly transport calcium against a gradient of Na(+) ions. Na(+)/Ca(2+) exchangers are divided into three groups based upon substrate specificity: Na(+)/Ca(2+) exchangers (NCX), Na(+)/Ca(2+)/K(+) exchangers (NCKX), and Ca(2+)/cation exchangers (NCLX). In mammals, there are three NCX genes, five NCKX genes, and a single NCLX gene. The genome of the nematode Caenorhabditis elegans contains 10 Na(+)/Ca(2+) exchanger genes: three NCX, five NCLX, and two NCKX genes. In a previous study, we characterized the structural and taxonomic specializations within the family of Na(+)/Ca(2+) exchangers across the phylum Nematoda and observed a complex picture of Na(+)/Ca(2+) exchanger evolution across diverse nematode species. We noted multiple cases of putative gene gain and loss and, most surprisingly, did not detect members of the NCLX type of exchangers within subsets of nematode species. In this commentary, we discuss these findings and speculate on the functional outcomes and physiology of these observations. Our data highlight the importance of studying diverse systems in order to get a deeper understanding of the evolution and regulation of Ca(2+) signaling critical for animal function.

Yang NC et al. (2019) Int J Mol Sci "The Lifespan Extension Ability of Nicotinic Acid Depends on Whether the Intracellular NAD+ Level Is Lower than the Sirtuin-Saturating Concentrations."

Yang NC, Lee I, Cho YH

[Int J Mol Sci, 2019]

Calorie restriction can extend lifespan by increasing intracellular nicotinamide adenine dinucleotide (NAD⁺), thereby upregulating the activity of sirtuins (<i>Caenorhabditis elegans</i> Sir-2.1; human SIRT1). Nicotinic acid (NA) can be metabolized to NAD⁺; however, the calorie restriction mimetic (CRM) potential of NA is unclear. This study explored the ability and mechanism of NA to extend the lifespan of human Hs68 cells and C. elegans. We found that NA can efficiently increase the intracellular NAD⁺ levels in Hs68 cells and <i>C. elegans</i>; however, NA was only able to extend the lifespan of <i>C. elegans</i>. The steady-state NAD⁺ level in <i>C. elegans</i> was approximately 55 M. When intracellular NAD⁺ was increased by a mutation of pme-1 (poly (ADP-ribose) metabolism enzyme 1) or by pretreatment with NAD⁺ in the medium, the lifespan extension ability of NA disappeared. Additionally, the saturating concentration of NAD⁺ required by SIRT1 was approximately 200 M; however, the steady-state concentration of NAD⁺ in Hs68 cells reached up to 460 M. These results demonstrate that the lifespan extension ability of NA depends on whether the intracellular level of NAD⁺ is lower than the sirtuin-saturating concentration in Hs68 cells and in <i>C. elegans</i>. Thus, the CRM potential of NA should be limited to individuals with lower intracellular NAD⁺.

He C et al. (2014) PLoS One "Analysis of the Na+/Ca2+ exchanger gene family within the phylum Nematoda."

He C, O'Halloran DM

[PLoS One, 2014]

Na+/Ca2+ exchangers are low affinity, high capacity transporters that rapidly transport calcium at the plasma membrane, mitochondrion, endoplasmic (and sarcoplasmic) reticulum, and the nucleus. Na+/Ca2+ exchangers are widely expressed in diverse cell types where they contribute homeostatic balance to calcium levels. In animals, Na+/Ca2+ exchangers are divided into three groups based upon stoichiometry: Na+/Ca2+ exchangers (NCX), Na+/Ca2+/K+ exchangers (NCKX), and Ca2+/Cation exchangers (CCX). In mammals there are three NCX genes, five NCKX genes and one CCX (NCLX) gene. The genome of the nematode Caenorhabditis elegans contains ten Na+/Ca2+ exchanger genes: three NCX; five CCX; and two NCKX genes. Here we set out to characterize structural and taxonomic specializations within the family of Na+/Ca2+ exchangers across the phylum Nematoda. In this analysis we identify Na+/Ca2+ exchanger genes from twelve species of nematodes and reconstruct their phylogenetic and evolutionary relationships. The most notable feature of the resulting phylogenies was the heterogeneous evolution observed within exchanger subtypes. Specifically, in the case of the CCX exchangers we did not detect members of this class in three Clade III nematodes. Within the Caenorhabditis and Pristionchus lineages we identify between three and five CCX representatives, whereas in other Clade V and also Clade IV nematode taxa we only observed a single CCX gene in each species, and in the Clade III nematode taxa that we sampled we identify NCX and NCKX encoding genes but no evidence of CCX representatives using our mining approach. We also provided re-annotation for predicted CCX gene structures from Heterorhabditis bacteriophora and Caenorhabditis japonica by RT-PCR and sequencing. Together, these findings reveal a complex picture of Na+/Ca2+ transporters in nematodes that suggest an incongruent evolutionary history of proteins that provide central control of calcium dynamics.

Kaplan JM (1993) Curr Biol "Neurodegeneration: great minds think alike."

Kaplan JM

[Curr Biol, 1993]

The recently cloned gene for the rat epithelial Na+ channel turns out to be related to nematode genes, mutations in which affect mechanosensation and cause neurodegeneration.