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Shuai Huang, Aruna Balgi, Yaping Pan, Meng Li, Xiaoran Zhang, Lilin Du, Ming Zhou, Michel Roberge, Xin Li
Nucleotide-binding leucine-rich repeat (NLR) proteins serve as immune receptors in both plants and animals. To identify components required for NLR-mediated immunity, we designed and carried out a chemical genetics screen to search for small molecules that can alter immune responses in Arabidopsis thaliana. From 13 600 compounds, we identified Ro 8-4304 that was able to specifically suppress the severe autoimmune phenotypes of chs3-2D (chilling sensitive 3, 2D), including the arrested growth morphology and heightened PR (Pathogenesis Related) gene expression...
December 5, 2016: Molecular Plant
Pengfei Fu, YueTao Wen, Yan Xiong, Yanke Zhang, Haiyang Zhang, Yanfeng Xie, Quanhong Shi
E3 ubiquitin ligases are important protein-modifying enzymes involved in the pathogenesis of a variety of neurodegenerative diseases. F-box and leucine-rich repeat protein 20 (FBXL20), an E3 ubiquitin ligase widely expressed in the central nervous system, plays an important role in the ubiquitin-dependent degradation of regulating synaptic membrane exocytosis 1 (RIM1), which is an important factor in the release of synaptic vesicles. FBXL20 has been associated with a variety of neurodegenerative diseases; thus, we hypothesized that FBXL20 is involved in the development of epilepsy...
November 2016: Neurochemical Research
Aparna Ramachandran, Divya Nandakumar, Aishwarya P Deshpande, Thomas P Lucas, Ramanagouda R-Bhojappa, Guo-Qing Tang, Kevin Raney, Y Whitney Yin, Smita S Patel
Primases use single-stranded (ss) DNAs as templates to synthesize short oligoribonucleotide primers that initiate lagging strand DNA synthesis or reprime DNA synthesis after replication fork collapse, but the origin of this activity in the mitochondria remains unclear. Herein, we show that the Saccharomyces cerevisiae mitochondrial RNA polymerase (Rpo41) and its transcription factor (Mtf1) is an efficient primase that initiates DNA synthesis on ssDNA coated with the yeast mitochondrial ssDNA-binding protein, Rim1...
August 5, 2016: Journal of Biological Chemistry
Alasdair N Warwick, Fatima Shawkat, Andrew J Lotery
BACKGROUND: Autosomal dominant cone-rod dystrophy 7 (CORD7) has been previously associated with the RIM1 c.2459G>A (Arg820His) mutation. Cystoid macular oedema (CMO) is a rare feature of CORD and has not been described in CORD7. We report a patient who was heterozygous for the RIM1 mutation with bilateral CMO and who manifested a retinitis pigmentosa phenotype. MATERIALS AND METHODS: The patient's medical notes were retrospectively reviewed over an 18-month period...
May 13, 2016: Ophthalmic Genetics
Mahdokht Kohansal-Nodehi, John Je Chua, Henning Urlaub, Reinhard Jahn, Dominika Czernik
Neurotransmitter release is mediated by the fast, calcium-triggered fusion of synaptic vesicles with the presynaptic plasma membrane, followed by endocytosis and recycling of the membrane of synaptic vesicles. While many of the proteins governing these processes are known, their regulation is only beginning to be understood. Here we have applied quantitative phosphoproteomics to identify changes in phosphorylation status of presynaptic proteins in resting and stimulated nerve terminals isolated from the brains of Wistar rats...
2016: ELife
Boris Zybailov, Kuppan Gokulan, Jadon Wiese, Ramanagouda Ramanagoudr-Bhojappa, Alicia K Byrd, Galina Glazko, Mihir Jaiswal, Samuel Mackintosh, Kottayil I Varughese, Kevin D Raney
Defining protein-protein contacts is a challenging problem and cross-linking is a promising solution. Here, we present a case of mitochondrial single strand binding protein Rim1 and helicase Pif1, an interaction first observed in immuno-affinity pull-down from yeast cells using Pif1 bait. We found that only the short succinimidyl-diazirine cross-linker or formaldehyde captured the interaction between recombinant Rim1 and Pif1. In addition, Pif1 needed to be stripped of its N-terminal and C-terminal domains, and Rim1's C-terminus needed to be modified for the cross-linked product to become visible...
November 2015: Journal of Proteomics & Bioinformatics
Eugenia Sanchez-Sandoval, Corina Diaz-Quezada, Gilberto Velazquez, Luis F Arroyo-Navarro, Norineli Almanza-Martinez, Carlos H Trasviña-Arenas, Luis G Brieba
Three proteins phylogenetically grouped with proteins from the T7 replisome localize to yeast mitochondria: DNA polymerase γ (Mip1), mitochondrial RNA polymerase (Rpo41), and a single-stranded binding protein (Rim1). Human and T7 bacteriophage RNA polymerases synthesize primers for their corresponding DNA polymerases. In contrast, DNA replication in yeast mitochondria is explained by two models: a transcription-dependent model in which Rpo41 primes Mip1 and a model in which double stranded breaks create free 3' OHs that are extended by Mip1...
September 2015: Mitochondrion
Patricia Gaspar, Xavier Nicol, Nicolas Narboux-Nême, Alexandra Rebsam
Numerous neurotransmitters have been implicated in neurodevelopmental processes. In addition, developing neurons show an abundance of vesicles in the growth cones, and express proteins of the SNARE complex early on. This has led to propose a role for vesicular fusion machinery in axonal growth and synapse formation. However, as the molecular machinery of vesicular fusion started to unveil, and knockouts for the major proteins of this complex were generated, it came as a surprise that none of these proteins was essential for the construction of brain architecture, although they were crucial for vital functions of the organism, leading to early mortality of exocytosis mutants...
2015: Biologie Aujourd'hui
Yiguo Shen, Woo-Ping Ge, Yulong Li, Arisa Hirano, Hsien-Yang Lee, Astrid Rohlmann, Markus Missler, Richard W Tsien, Lily Yeh Jan, Ying-Hui Fu, Louis J Ptáček
Paroxysmal nonkinesigenic dyskinesia (PNKD) is an autosomal dominant episodic movement disorder precipitated by coffee, alcohol, and stress. We previously identified the causative gene but the function of the encoded protein remains unknown. We also generated a PNKD mouse model that revealed dysregulated dopamine signaling in vivo. Here, we show that PNKD interacts with synaptic active zone proteins Rab3-interacting molecule (RIM)1 and RIM2, localizes to synapses, and modulates neurotransmitter release. Overexpressed PNKD protein suppresses release, and mutant PNKD protein is less effective than wild-type at inhibiting exocytosis...
March 10, 2015: Proceedings of the National Academy of Sciences of the United States of America
S M Samir, H A Malek
Osteoporosis is a common health problem. The endocannabinoid pathway has been implicated as an important regulator of bone turnover. Rimonabant is a potent cannabinoid receptor1 (CB1) receptor antagonist with wide therapeutic use as an antiobesity drug that has been withdrawn due to side effects in the form of depression and suicidal attacks. This study investigated whether glucocorticoid induced bone loss is linked to CB1 signaling and whether modulation of CB1 function affects the deleterious effects of glucocorticoid treatment on bone remodeling in rats...
October 2014: Journal of Physiology and Pharmacology: An Official Journal of the Polish Physiological Society
Aparna Rajagopalan, Nadine Schweizer, Karin Nordenankar, Sultana Nilufar Jahan, Lina Emilsson, Asa Wallén-Mackenzie
Presynaptic proteins orchestrate an intricate interplay of dynamic interactions in order to regulate quantal exocytosis of transmitter-filled vesicles, and their dysregulation might cause neurological and neuropsychiatric dysfunction. Mice carrying a spatio-temporal restriction in the expression of the Vesicular glutamate transporter 2 (Vglut2; aka Slc17a6) in the cortex, amygdala and hippocampal subiculum from the third postnatal week show a strong anxiolytic phenotype and certain behavioural correlates of schizophrenia...
August 19, 2014: Synapse
Rizwana Anjum, Hiresh Ayoubian, Frank Schmitz
Scaffold proteins organize pre- and postsynaptic compartments and align pre- and postsynaptic events. Cask is a multi-domain scaffold protein essential for brain synaptic functions. Caskin1 is a recently discovered, brain-specific Cask-interacting multi-domain protein of unknown function. In the present study, we determined the localization of these scaffold proteins in the bovine retina. The retina contains tonically active ribbon synapses and conventional synapses. We found Cask highly enriched in virtually all retinal synapses...
September 2014: Molecular and Cellular Neurosciences
Alan Jung Park, Robbert Havekes, Jennifer H K Choi, Vincent Luczak, Ting Nie, Ted Huang, Ted Abel
Protein kinase A (PKA) and other signaling molecules are spatially restricted within neurons by A-kinase anchoring proteins (AKAPs). Although studies on compartmentalized PKA signaling have focused on postsynaptic mechanisms, presynaptically anchored PKA may contribute to synaptic plasticity and memory because PKA also regulates presynaptic transmitter release. Here, we examine this issue using genetic and pharmacological application of Ht31, a PKA anchoring disrupting peptide. At the hippocampal Schaffer collateral CA3-CA1 synapse, Ht31 treatment elicits a rapid decay of synaptic responses to repetitive stimuli, indicating a fast depletion of the readily releasable pool of synaptic vesicles...
October 2014: Neurobiology of Learning and Memory
Jorge Ramírez-Franco, David Bartolomé-Martín, Beatris Alonso, Magdalena Torres, José Sánchez-Prieto
Cannabinoid receptors are the most abundant G protein-coupled receptors in the brain and they mediate retrograde short-term inhibition of neurotransmitter release, as well as long-term depression of synaptic transmission at many excitatory synapses. The induction of presynaptically silent synapses is a means of modulating synaptic strength, which is important for synaptic plasticity. Persistent activation of cannabinoid type 1 receptors (CB1Rs) mutes GABAergic terminals, although it is unclear if CB1Rs can also induce silencing at glutamatergic synapses...
2014: PloS One
Fatima Girach, Tim J Craig, Daniel L Rocca, Jeremy M Henley
The rapid, activity-dependent quantal presynaptic release of neurotransmitter is vital for brain function. The complex process of vesicle priming, fusion, and retrieval is very precisely controlled and requires the spatiotemporal coordination of multiple protein-protein interactions. Here, we show that posttranslational modification of the active zone protein Rab3-interacting molecule 1α (RIM1α) by the small ubiquitin-like modifier 1 (SUMO-1) functions as a molecular switch to direct these interactions and is essential for fast synaptic vesicle exocytosis...
December 12, 2013: Cell Reports
Xin Bi, Yibao Jin, Xiang Gao, Feng Liu, Dan Gao, Yuyang Jiang, Hongxia Liu
Pokemon is a transcription regulator involved in embryonic development, cellular differentiation and oncogenesis. It is aberrantly overexpressed in multiple human cancers including Hepatocellular carcinoma (HCC) and is considered as a promising biomarker for HCC. In this work, the isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics strategy was used to investigate the proteomic profile associated with Pokemon in human HCC cell line QGY7703 and human hepatocyte line HL7702...
2013: European Journal of Mass Spectrometry
Süzel Horellou, Olivier Pascual, Antoine Triller, Serge Marty
How the number of docked vesicles is regulated is still unclear. Following chronic activity blockade the number of docked vesicles increases, providing a model through which to address this issue. We tested the hypotheses that the number of docked vesicles is regulated with the size of the terminal, and by the level of Rab3-interacting molecule 1/2 (RIM1/2). We immobilized mouse hippocampal slice cultures by high-pressure freezing after 3 days of tetrodotoxin treatment and analysed them by electron microscopy...
January 2014: European Journal of Neuroscience
Jorge Ramírez-Franco, Beatris Alonso, David Bartolomé-Martín, José Sánchez-Prieto, Magdalena Torres
BACKGROUND: In terms of vesicular recycling, synaptic efficiency is a key determinant of the fidelity of synaptic transmission. The ability of a presynaptic terminal to reuse its vesicular content is thought to be a signature of synaptic maturity and this process depends on the activity of several proteins that govern exo/endocytosis. Upon stimulation, individual terminals in networks of cultured cerebellar granule neurons exhibit heterogeneous exocytic responses, which reflect the distinct states of maturity and plasticity intrinsic to individual synaptic terminals...
2013: BMC Neuroscience
Jose J Ferrero, Ana M Alvarez, Jorge Ramírez-Franco, María C Godino, David Bartolomé-Martín, Carolina Aguado, Magdalena Torres, Rafael Luján, Francisco Ciruela, José Sánchez-Prieto
The adenylyl cyclase activator forskolin facilitates synaptic transmission presynaptically via cAMP-dependent protein kinase (PKA). In addition, cAMP also increases glutamate release via PKA-independent mechanisms, although the downstream presynaptic targets remain largely unknown. Here, we describe the isolation of a PKA-independent component of glutamate release in cerebrocortical nerve terminals after blocking Na(+) channels with tetrodotoxin. We found that 8-pCPT-2'-O-Me-cAMP, a specific activator of the exchange protein directly activated by cAMP (Epac), mimicked and occluded forskolin-induced potentiation of glutamate release...
October 25, 2013: Journal of Biological Chemistry
Michael Kintscher, Christian Wozny, Friedrich W Johenning, Dietmar Schmitz, Jörg Breustedt
The presynaptic terminals of synaptic connections are composed of a complex network of interacting proteins that collectively ensure proper synaptic transmission and plasticity characteristics. The key components of this network are the members of the RIM protein family. Here we show that RIM1α can influence short-term plasticity at cerebellar parallel-fibre synapses. We demonstrate that the loss of a single RIM isoform, RIM1α, leads to reduced calcium influx in cerebellar granule cell terminals, decreased release probability and consequently an enhanced short-term facilitation...
2013: Nature Communications
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