The steroid hormone progesterone (P4) regulates multiple aspects of reproductive and metabolic physiology. Classical P4 signaling operates through nuclear receptors that regulate transcription. In addition, signals membrane (mPRs) in a rapid nongenomic modality. Despite the established physiological importance signaling, details its signal transduction cascade remain elusive. Here, using Xenopus oocyte maturation as well-established readout we identify lipid hydrolase ABHD2 (α/β...

The minimal machinery mediating store operated Ca2+ entry (SOCE) include an ER sensor -STIM1- and a plasma membrane (PM) Ca2+-selective channel Orai1. Here we quantitatively dissect Orai1 trafficking dynamics show that recycles rapidly at the PM (Kex ≃ 0.1 min-1), with ∼40% of total pool localizing to steady state. A subset intracellular localizes sub-plasmalemal compartment. Store depletion is coupled enrichment in STIM1-dependent fashion. This due trapping into cortical STIM1 clusters...

The steroid hormone progesterone (P4) mediates many physiological processes through either nuclear receptors that modulate gene expression or membrane P4 (mPRs) mediate nongenomic signaling. mPR signaling remains poorly understood. Here we show the topology of mPRβ is similar to adiponectin and opposite G-protein-coupled (GPCRs). Using Xenopus oocyte meiosis as a well-established readout signaling, demonstrate requires adaptor protein APPL1 kinase Akt2. We further induces clathrin-dependent...

Vertebrate oocytes arrest at prophase of meiosis I due to high levels cAMP and PKA activity. In Xenopus progesterone is believed release meiotic by inhibiting adenylate cyclase, lowering levels, repressing protein kinase A (PKA). However the exact timing extent decrease unclear with conflicting reports in literature. Using various vivo reporters for single cell level real time, we fail detect any significant changes or response progesterone. More interestingly, there was no correlation...

Ca(2+)-activated Cl(-) channels (CaCCs) play important physiological functions in epithelia and other tissues. In frog oocytes the CaCC Ano1 regulates resting membrane potential block to polyspermy. Here, we show that expression increases oocyte surface, revealing a novel function for regulating cell morphology. Confocal imaging shows microvilli length, which requires ERM-protein-dependent linkage cytoskeleton. A dominant-negative form of ERM protein moesin precludes Ano1-dependent increase...

Vertebrate oocytes are naturally arrested at prophase of meiosis I for sustained periods time before resuming in a process called oocyte maturation that prepares the egg fertilization. Members constitutively active GPR3/6/12 family G-protein coupled receptors represent important mediators meiotic arrest. In frog GPR3/12 homolog GPRx (renamed GPR185) has been shown to sustain arrest by increasing intracellular cAMP levels through GαSβγ. Here we show is enriched cell membrane (~80%), recycles...

Agonist-dependent Ca2+ mobilization results in store depletion and Store-Operated Calcium Entry (SOCE), which is spatially restricted to microdomains defined by cortical ER - plasma membrane contact sites (MCS). However, some Ca2+-dependent effectors that localize away from SOCE microdomains, are activated downstream of mechanisms remain obscure. One mechanism proposed initially acinar cells termed tunneling, mediates the uptake flowing through into followed release at distal IP3 receptors....

Ca2+ signaling is ubiquitous and mediates various cellular functions encoded in its spatial, temporal, amplitude features. Here, we investigate the role of store-operated entry (SOCE) regulating temporal dynamics signals Xenopus oocytes, which can be either oscillatory or tonic. Oscillatory release from intracellular stores typically observed at physiological agonist concentration. When leads to store depletion, this triggers activation SOCE that translates into a low-amplitude tonic signal....

The steroid hormone progesterone (P4) mediates many physiological processes through either nuclear receptors that modulate gene expression or membrane P4 (mPRs) mediate nongenomic signaling. mPR signaling remains poorly understood. Here we show the topology of mPRβ is similar to adiponectin and opposite G-protein-coupled (GPCRs). Using Xenopus oocyte meiosis as a well-established readout signaling, demonstrate requires adaptor protein APPL1 kinase Akt2. We further induces clathrin-dependent...

ABSTRACT Progesterone mediates its physiological functions through activation of both transcription-coupled nuclear receptors and seven-pass-transmembrane progesterone (mPRs), which transduce the rapid non-genomic actions by coupling to various signaling modules. However, immediate mechanisms action downstream mPRs remain in question. Herein, we use an untargeted quantitative proteomics approach identify mPR interactors better define signaling. Surprisingly, very-low-density lipoprotein...

The steroid hormone progesterone (P4) regulates multiple aspects of reproductive and metabolic physiology. Classical P4 signaling operates through nuclear receptors that regulate transcription. In addition, signals membrane (mPRs) in a rapid nongenomic modality. Despite the established physiological importance signaling, its detailed signal transduction remains elusive. Here, using Xenopus oocyte maturation as well-established readout we identify lipid hydrolase ABHD2 (α/β domain-containing...

The TRP gene family encodes primarily cation non-selective, Ca2+ permeant channels that are involved in a dizzying array of sensory mechanisms. Two this large TRPV5 and TRPV6 highly selective expressed epithelia where they important uptake. TRPV5/6 constitutively active, yet the mechanisms regulating their activation native tissue remains elusive. Here we functionally characterize Xenopus homolog. xTRPV6 is oocyte channel to divalents including , displays high permeability Mg2+ . does not...

Abstract Store-operated Ca 2+ entry (SOCE) has been shown to be important for breast cancer metastasis in xenograft mouse models. The ER sensor STIM1 and Orai plasma membrane channels molecularly mediate SOCE. Here we investigate the role of microRNA machinery regulating expression. We show that expression is regulated post-transcriptionally by miRNA identify miR-223 miR-150 as regulators luminal non-aggressive MCF7 cell line. In contrast, more aggressive basal triple-negative MDA-MB-231...

The steroid hormone progesterone (P4) regulates multiple aspects of reproductive and metabolic physiology. Classical P4 signaling operates through nuclear receptors that regulate transcription. In addition, signals membrane (mPRs) in a rapid nongenomic modality. Despite the established physiological importance signaling, details its signal transduction cascade remain elusive. Here, using Xenopus oocyte maturation as well- readout we identify lipid hydrolase ABHD2 (α/β domain-containing...

The steroid hormone progesterone (P4) regulates multiple aspects of reproductive and metabolic physiology. Classical P4 signaling operates through nuclear receptors that regulate transcription. In addition, signals membrane (mPRs) in a rapid nongenomic modality. Despite the established physiological importance signaling, details its signal transduction cascade remain elusive. Here, using Xenopus oocyte maturation as well- readout we identify lipid hydrolase ABHD2 (α/β domain-containing...

Abstract Regulation of Ca 2+ signaling is critical for the progression cell division, especially during meiosis to prepare egg fertilization. The primary influx pathway in oocytes Store-Operated Entry (SOCE). SOCE tightly regulated meiosis, including internalization channel, Orai1. Orai1 a four-pass membrane protein with cytosolic N- and C-termini. requires caveolin binding motif (CBM) N-terminus as well C-terminal domain. However, molecular determinant endocytosis C-terminus are not known....

Store-operated calcium entry (SOCE) is a ubiquitous Ca2+ influx pathway essential for many physiological functions. Dysregulation of SOCE causes disruption in homeostasis leading to cellular pathology several diseases. Orai1, key regulator SOCE, constitutively recycles at steady state the frog oocyte and internalizes into an intracellular vesicular compartments during meiosis, inactivation SOCE. Previous data showed role Orai1 C-terminus its internalization meiosis. However, minimal region...

The endoplasmic reticulum (ER) functions as a storehouse for intracellular calcium. STIM1, calcium sensor, localizes mostly to the ER membrane. Following Ca2+ store depletion, STIM1 forms puncta that localize cortical and bind Orai1, plasma membrane channel, allow influx. This is predominant pathway influx in non-excitable cells referred Store-Operated Calcium Entry (SOCE). Mutations Orai1 cause severe combined immunodeficiencies are linked several of cancers. Tight regulation levels crucial...

Store-operated calcium entry (SOCE) is a ubiquitous Ca 2+ influx pathway essential for many physiological functions and failure to maintain normal homeostasis one of the leading causes cellular dysfunction in wide variety pathological conditions. Orai1, key regulator SOCE, constitutively recycles at steady state frog oocyte internalizes into intracellular vesicular compartments during meiosis, inactivation SOCE. Such mechanism provides proper regulation signaling preparation fertilization...

Abstract The steroid hormone progesterone (P4) regulates multiple aspects of reproductive and metabolic physiology. Classical P4 signaling operates through nuclear receptors that regulate transcription. In addition, signals membrane (mPRs) in a rapid nongenomic modality. Despite the established physiological importance signaling, details its signal transduction cascade remain elusive. Here, using Xenopus oocyte maturation as well- readout we identify lipid hydrolase ABHD2 (α/β...

Stromal interaction molecule 1 (STIM1), the ER calcium sensor, couples to Orai1, channel, and mediates store-operated entry (SOCE). STIM1 localizes membrane. Following Ca²+ store depletion, forms puncta that localize cortical binds Orai1 allow influx. The egg's competency activate at fertilization is dependent on its ability generate a fertilization-specific Ca2+ transient. This achieved through remodeling of signaling machinery during oocyte maturation. Oocyte maturation driven by kinase...