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Barbara Mandriani, Stefano Castellana, Carmela Rinaldi, Marta Manzoni, Santina Venuto, Eva Rodriguez-Aznar, Juan Galceran, M Angela Nieto, Giuseppe Borsani, Eugenio Monti, Tommaso Mazza, Giuseppe Merla, Lucia Micale
To orchestrate the genomic response to cellular stress signals, p53 recognizes and binds to DNA containing specific and well-characterized p53-responsive elements (REs). Differences in RE sequences can strongly affect the p53 transactivation capacity and occur even between closely related species. Therefore, the identification and characterization of a species-specific p53 Binding sistes (BS) consensus sequence and of the associated target genes may help to provide new insights into the evolution of the p53 regulatory networks across different species...
2016: Scientific Reports
Ming-Dong Zhang, Swapnali Barde, Edit Szodorai, Anna Josephson, Nicholas Mitsios, Masahiko Watanabe, Johannes Attems, Gert Lubec, Gábor G Kovács, Mathias Uhlén, Jan Mulder, Tibor Harkany, Tomas Hökfelt
Neuronal calcium-binding protein 1 and -2 (NECAB1/2) localize to multiple excitatory neuron populations in the mouse spinal cord. Here, we analyzed rat and human spinal cord, combining in situ hybridization and immunohistochemistry, complementing newly collated data on mouse spinal cord for direct comparisons. Necab1/2 mRNA transcripts showed complementary distribution in rodent's spinal cord. Multiple-labeling fluorescence histochemistry with neuronal phenotypic markers localized NECAB1 to a dense fiber plexus in the dorsal horn, to neurons mainly in superficial layers and to commissural interneurons in both rodent species...
September 2016: Brain Structure & Function
David C Schöndorf, Massimo Aureli, Fiona E McAllister, Christopher J Hindley, Florian Mayer, Benjamin Schmid, S Pablo Sardi, Manuela Valsecchi, Susanna Hoffmann, Lukas Kristoffer Schwarz, Ulrike Hedrich, Daniela Berg, Lamya S Shihabuddin, Jing Hu, Jan Pruszak, Steven P Gygi, Sandro Sonnino, Thomas Gasser, Michela Deleidi
Mutations in the acid β-glucocerebrosidase (GBA1) gene, responsible for the lysosomal storage disorder Gaucher's disease (GD), are the strongest genetic risk factor for Parkinson's disease (PD) known to date. Here we generate induced pluripotent stem cells from subjects with GD and PD harbouring GBA1 mutations, and differentiate them into midbrain dopaminergic neurons followed by enrichment using fluorescence-activated cell sorting. Neurons show a reduction in glucocerebrosidase activity and protein levels, increase in glucosylceramide and α-synuclein levels as well as autophagic and lysosomal defects...
June 6, 2014: Nature Communications
Ming-Dong Zhang, Giuseppe Tortoriello, Brian Hsueh, Raju Tomer, Li Ye, Nicholas Mitsios, Lotta Borgius, Gunnar Grant, Ole Kiehn, Masahiko Watanabe, Mathias Uhlén, Jan Mulder, Karl Deisseroth, Tibor Harkany, Tomas G M Hökfelt
Neuronal calcium (Ca(2+))-binding proteins 1 and 2 (NECAB1/2) are members of the phylogenetically conserved EF-hand Ca(2+)-binding protein superfamily. To date, NECABs have been explored only to a limited extent and, so far, not at all at the spinal level. Here, we describe the distribution, phenotype, and nerve injury-induced regulation of NECAB1/NECAB2 in mouse dorsal root ganglia (DRGs) and spinal cord. In DRGs, NECAB1/2 are expressed in around 70% of mainly small- and medium-sized neurons. Many colocalize with calcitonin gene-related peptide and isolectin B4, and thus represent nociceptors...
March 25, 2014: Proceedings of the National Academy of Sciences of the United States of America
Yasunari Sakai, Chad A Shaw, Brian C Dawson, Diana V Dugas, Zaina Al-Mohtaseb, David E Hill, Huda Y Zoghbi
To uncover shared pathogenic mechanisms among the highly heterogeneous autism spectrum disorders (ASDs), we developed a protein interaction network that identified hundreds of new interactions among proteins encoded by ASD-associated genes. We discovered unexpectedly high connectivity between SHANK and TSC1, previously implicated in syndromic autism, suggesting that common molecular pathways underlie autistic phenotypes in distinct syndromes. ASD patients were more likely to harbor copy number variations that encompass network genes than were control subjects...
June 8, 2011: Science Translational Medicine
Simon Keuerleber, Ingrid Gsandtner, Michael Freissmuth
The extended carboxyl terminus of the A(2A)-adenosine receptor is known to engage several proteins other than those canonically involved in signalling by GPCRs (i.e., G proteins, G protein-coupled receptor kinases/GRKs, arrestins). The list includes the deubiquinating enzyme USP4, α-actinin, the guanine nucleotide exchange factor for ARF6 ARNO, translin-X-associated protein, calmodulin, the neuronal calcium binding protein NECAB2 and the synapse associated protein SAP102. However, if the fate of the A(2A)-receptor is taken into account - from its birthplace in the endoplasmic reticulum to its presumed site of disposal in the lysosome, it is evident that many more proteins must interact with the A(2A)-adenosine receptor...
May 2011: Biochimica et Biophysica Acta
Laia Canela, Víctor Fernández-Dueñas, Catarina Albergaria, Masahiko Watanabe, Carme Lluís, Josefa Mallol, Enric I Canela, Rafael Franco, Rafael Luján, Francisco Ciruela
Metabotropic glutamate (mGlu) receptors mediate in part the CNS effects of glutamate. These receptors interact with a large array of intracellular proteins in which the final role is to regulate receptor function. Here, using co-immunoprecipitation and pull-down experiments we showed a close and specific interaction between mGlu(5) receptor and NECAB2 in both transfected human embryonic kidney cells and rat hippocampus. Interestingly, in pull-down experiments increasing concentrations of calcium drastically reduced the ability of these two proteins to interact, suggesting that NECAB2 binds to mGlu(5) receptor in a calcium-regulated manner...
October 2009: Journal of Neurochemistry
Laia Canela, Rafael Luján, Carme Lluís, Javier Burgueño, Josefa Mallol, Enric I Canela, Rafael Franco, Francisco Ciruela
Heptaspanning membrane also known as G protein-coupled receptors (GPCR) do interact with a variety of intracellular proteins whose function is regulate receptor traffic and/or signaling. Using a yeast two-hybrid screen, NECAB2, a neuronal calcium binding protein, was identified as a binding partner for the adenosine A(2A) receptor (A(2A)R) interacting with its C-terminal domain. Co-localization, co-immunoprecipitation and pull-down experiments showed a close and specific interaction between A(2A)R and NECAB2 in both transfected HEK-293 cells and also in rat striatum...
September 2007: Molecular and Cellular Neurosciences
Hai Wu, Dan Li, Yuxi Shan, Bo Wan, Saiyin Hexige, Jinhu Guo, Chaoqun Wu, Long Yu
Human EFCBP/NECAB family consists of important participants in neuronal calcium signaling, including EFCBP1/NECAB1, EFCBP2/NECAB2 and EFCBP3/NECAB3. In the present study, we identified the full-length 5229 bp EFCBP1 cDNA which was not described before. Human EFCBP1 encodes a 351 amino acid protein containing two EF-hand motifs and an antibiotic biosynthesis monooxygenase (ABM) domain, sharing 49.9 and 56.8% global homology with human EFCBP2 and EFCBP3. Northern hybridization revealed that EFCBP1 is specifically expressed in brain and its abundance varies in different brain regions...
February 2007: DNA Sequence: the Journal of DNA Sequencing and Mapping
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