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Journal of Cell Science

Robert W Figliozzi, Feng Chen, Shaochung V Hsia
Thyroid hormones (TH or T3) and their DNA binding nuclear receptors (TRs), direct transcriptional regulation in diverse ways depending on the host cell environment and specific promoter characteristics of TH sensitive genes. This study sought to elucidate the impact on transcriptional repression of nucleotide sequence/orientation within TR binding sites, TR elements, (TREs) of TH sensitive promoters, to better understand ligand dependent transcriptional repression of wild-type promoters. Computational analysis of the HSV-1 thymidine kinase (TK) gene TRE bound by TR and RXR revealed a single TRE point mutation sufficient to reverse the TRE orientation...
September 15, 2017: Journal of Cell Science
Michael V Keebler, Colin W Taylor
The building blocks for intracellular Ca(2+) signals evoked by inositol 1,4,5-trisphosphate receptors (IP3Rs) are Ca(2+) puffs, transient focal increases in Ca(2+) concentration that reflect the opening of a small clusters of IP3Rs. We use total internal reflection fluorescence microscopy and automated analyses to detect Ca(2+) puffs in human embryonic kidney 293 cells evoked by photolysis of caged-IP3 or activation of endogenous muscarinic receptors with carbachol. Ca(2+) puffs evoked by carbachol initiated at an estimated 65 ± 7 sites/cell, and the sites remained immobile for many minutes...
September 11, 2017: Journal of Cell Science
Jan de Vries, Sven B Gould
Plastids in plants and algae evolved from the endosymbiotic integration of a cyanobacterium by a heterotrophic eukaryote. New plastids can only emerge through fission; thus, the synchronization of bacterial division with the cell cycle of the eukaryotic host was vital to the origin of phototrophic eukaryotes. Most of the sampled algae house a single plastid per cell and basal-branching relatives of polyplastidic lineages are all monoplastidic, as are some non-vascular plants during certain stages of their life cycle...
September 11, 2017: Journal of Cell Science
Anna Frappaolo, Stefano Sechi, Tadahiro Kumagai, Sarah Robinson, Roberta Fraschini, Angela Karimpour Ghahnavieh, Giorgio Belloni, Roberto Piergentili, Katherine H Tiemeyer, Michael Tiemeyer, Maria Grazia Giansanti
Congenital Disorders of Glycosylation (CDG) comprise a family of human multi-systemic diseases caused by recessive mutations in genes required for protein N-glycosylation. More than 100 distinct forms of CDGs have been identified and most of them cause severe neurological impairment. The Conserved Oligomeric Golgi (COG) complex mediates tethering of vesicles carrying glycosylation enzymes across the Golgi cisternae. Mutations affecting human COG1, COG2, COG4-COG8 cause monogenic forms of inherited, autosomal recessive, CDGs...
September 7, 2017: Journal of Cell Science
Yandong Lai, Jin Li, Xiuying Li, Chunbin Zou
E3 ubiquitin ligase recognizes its protein substrates via specific molecular signatures for ubiquitin proteasomal degradation. However, the role of acetylation/ deacetylation in the process of E3 ubiquitin ligase recognizing its protein substrate is not fully studied. Here we report that a tandem IK motif in protein arginine methyltransferase 1 (PRMT1) forms an acetyldegron to recruit ubiquitin E3 ligase SCF (Skp1-Cullin1-Fbox protein)-Fbxl17. PRMT1 is poly-ubiquitinated for proteasome degradation with a half-life of approximate 4 h in lung epithelial cells...
September 7, 2017: Journal of Cell Science
Linda Goodman, Marisa Zallocchi
The organism's perception of its surroundings depends on sensory systems and the highly specialized cilia present in the neurosensory cells. Here, we described the existence of an integrin alpha 8 (Itga8)/protocadherin-15a (Pcdh15a) ciliary complex in neuromast hair cells. Depletion of the complex via down-regulation or loss of function mutations leads to a dysregulation of cilia biogenesis and endocytosis. At the molecular level, removal of the complex blocks the access of Rab8a into the cilia as well as normal recruitment of ciliary cargo by centriolar satellites...
September 7, 2017: Journal of Cell Science
Valeria Trapani, Paolo Bonaldo, Diana Corallo
The notochord is a midline structure common to all chordate animals; it provides mechanical and signaling cues for the developing embryo. In vertebrates, the notochord plays key functions during embryogenesis, being a source of developmental signals that pattern the surrounding tissues. It is composed of a core of vacuolated cells surrounded by an epithelial-like sheath of cells that secrete a thick peri-notochordal basement membrane made of different extracellular matrix (ECM) proteins. The correct deposition and organization of the ECM is essential for proper notochord morphogenesis and function...
September 7, 2017: Journal of Cell Science
Elsa A Tungadi, Ami Ito, Tomomi Kiyomitsu, Gohta Goshima
Nonsense mutations in the ASPM gene have been most frequently identified among familial microcephaly patients. Depletion of the Drosophila orthologue causes spindle pole unfocusing during mitosis in multiple cell types. However, it remains unknown whether human ASPM has a similar function. Here, using CRISPR-based gene knockout (KO) and RNA interference combined with auxin-inducible degron, we show that ASPM functions in spindle pole organisation during mitotic metaphase redundantly with another microcephaly protein CDK5RAP2 (also called CEP215) in human tissue culture cells...
September 7, 2017: Journal of Cell Science
David Brough, Pablo Pelegrin, Walter Nickel
Extracellular proteins with important signalling roles in processes, such as inflammation and angiogenesis, are known to employ unconventional routes of protein secretion. Although mechanisms of unconventional protein secretion are beginning to emerge, the precise molecular details have remained elusive for the majority of cargo proteins secreted by unconventional means. Recent findings suggest that for two examples of unconventionally secreted proteins, interleukin 1β (IL-1β) and fibroblast growth factor 2 (FGF2), the common molecular principle of pore formation may be shared...
September 4, 2017: Journal of Cell Science
Cynthia Gomes, Seung Joon Lee, Amy S Gardiner, Terika Smith, Pabitra K Sahoo, Priyanka Patel, Elizabeth Thames, Reycel Rodriguez, Ross Taylor, Soonmoon Yoo, Tilman Heise, Amar N Kar, Nora Perrone-Bizzozero, Jeffery L Twiss
HuD protein has been shown to stabilize mRNAs with AU-rich elements (ARE) in their 3'UTRs, including Gap43 that has been linked to axon growth. HuD also binds to Neuritin (Nrn1), whose 3'UTR contains ARE sequences. Although Nrn1's 3'UTR was shown to have axonal localizing activity in embryonic hippocampal neurons, it was not active in adult DRG neurons. Here, we asked why the 3'UTR is not sufficient to localize Nrn1 mRNA into DRG axons. HuD overexpression increases the axonal localizing activity of Nrn1's 3'UTR in DRG...
September 4, 2017: Journal of Cell Science
Philipp P Prosseda, Na Luo, Biao Wang, Jorge A Alvarado, Yang Hu, Yang Sun
Lowe syndrome is a rare X-linked disorder characterized by bilateral congenital cataracts and glaucoma, mental retardation, and proximal renal tubular dysfunction. Mutations in OCRL1, an inositol polyphosphate 5-phosphatase that dephosphorylates PI(4,5)P2, cause Lowe syndrome. Previously we showed that OCRL localizes to the primary cilium, which has a distinct membrane phospholipid composition, but disruption of phosphoinositides in the ciliary membrane poorly understood. Here we demonstrate that cilia from Lowe syndrome patient fibroblasts exhibit increased levels of PI(4,5)P2 and decreased levels of PI4P...
September 4, 2017: Journal of Cell Science
Noelia Pastor-Cantizano, Cesar Bernat-Silvestre, María Jesús Marcote, Fernando Aniento
p24 proteins are key regulators of protein trafficking along the secretory pathway but very little is known about their functions in plants. A quadruple loss-of-function mutant affecting the p24 genes from the δ-1 subclass of the p24 delta subfamily (p24δ3δ4δ5δ6) showed alterations in the Golgi apparatus, suggesting that these p24 proteins play a role in the organization of the compartments of the early secretory pathway in Arabidopsis Loss of p24δ-1 proteins also induced the accumulation of the K/HDEL receptor ERD2 at the Golgi apparatus and increased secretion of the ER chaperone BiP, an HDEL ligand, probably due to an inhibition of COPI-dependent Golgi-to-ER transport of ERD2 and thus retrieval of K/HDEL ligands...
September 4, 2017: Journal of Cell Science
Olivia Susanto, Yvette W H Koh, Nick Morrice, Sergey Tumanov, Peter A Thomason, Matthew Nielson, Luke Tweedy, Andrew J Muinonen-Martin, Jurre J Kamphorst, Gillian M Mackay, Robert H Insall
Melanoma cells steer out of tumours using self-generated lysophosphatidic acid (LPA) gradients. The cells break down LPA, which is present at high levels around the tumours, creating a dynamic gradient that is low in the tumour and high outside. They then also migrate up this gradient, creating a complex and evolving outward chemotactic stimulus. Here we introduce a new assay for self-generated chemotaxis, and show that raising LPA levels causes a delay in migration rather than loss of chemotactic efficiency...
September 4, 2017: Journal of Cell Science
Floris Bosveld, Anna Ainslie, Yohanns Bellaïche
Centrosomes nucleate microtubules and are tightly coupled to the bipolar spindle to ensure genome integrity, cell division orientation and centrosome segregation. While the mechanisms of centrosome-dependent microtubule nucleation and bipolar spindle assembly have been the focus of numerous works, less is known on the mechanisms ensuring the centrosome-spindle coupling. The conserved NuMA protein (Mud in Drosophila) is best known for its role in spindle orientation. Here we analyzed the role of Mud and two of its interactors, Asp and Dynein, in the regulation of centrosome numbers in Drosophila epithelial cells...
September 1, 2017: Journal of Cell Science
Federica Valsecchi, Csaba Konrad, Marilena D'Aurelio, Lavoisier S Ramos-Espiritu, Anna Stepanova, Suzanne R Burstein, Alexander Galkin, Jordi Magranè, Anatoly Starkov, Jochen Buck, Lonny R Levin, Giovanni Manfredi
cAMP regulates a wide variety of physiological functions in mammals. This single second messenger can regulate multiple, seemingly disparate functions within independently regulated cell compartments. We previously identified one such compartment inside the matrix of the mitochondria, where soluble adenylyl cyclase (sAC) regulates oxidative phosphorylation (OXPHOS). We now show that sAC KO fibroblasts have a defect in OXPHOS activity and attempt to compensate for this defect by increasing OXPHOS proteins. Importantly, sAC KO cells also exhibit decreased probability of endoplasmic reticulum (ER) Ca(2+) release associated with diminished phosphorylation of the inositol 3-phosphate receptor...
September 1, 2017: Journal of Cell Science
Takafumi Ichikawa, Masahiro Kita, Tsubasa S Matsui, Ayaka Ichikawa Nagasato, Tomohiko Araki, Shian-Huey Chiang, Takuhito Sezaki, Yasuhisa Kimura, Kazumitsu Ueda, Shinji Deguchi, Alan R Saltiel, Noriyuki Kioka
Vinexin, CAP and ArgBP2 constitute an adaptor protein family called the vinexin (SORBS) family that is targeted to focal adhesions (FAs). Although numerous studies have focused on each of the SORBS proteins and partially elucidated their involvement in mechanotransduction, a comparative analysis of their function has not been well addressed. Here, we established mouse embryonic fibroblasts that individually expressed SORBS proteins and analyzed their functions in an identical cell context. Both vinexin α and CAP colocalized with vinculin at FAs and promoted the appearance of vinculin-rich FAs, whereas ArgBP2 colocalized with α-actinin at the proximal end of FAs and punctate structures on actin stress fibers (SFs), and induced paxillin-rich FAs...
September 1, 2017: Journal of Cell Science
Zhongji Liao, Ana Kasirer-Friede, Sanford J Shattil
αVβ3 is reported to promote angiogenesis in some model systems but not in others. Here we used optogenetics to study effects of αVβ3 interaction with the intracellular adapter, kindlin-2, on endothelial cell functions potentially relevant to angiogenesis. Since interaction of kindlin-2 with αVβ3 requires the C-terminal three residues of the β3 cytoplasmic tail (Arg-Gly-Thr; RGT), optogenetic probes LOVpep and ePDZ1 were fused to β3ΔRGT-GFP and mCherry-kindlin2, respectively, and expressed in β3-null microvascular endothelial cells...
September 1, 2017: Journal of Cell Science
Cristian De Gregorio, Ricardo Delgado, Andrés Ibacache, Jimena Sierralta, Andrés Couve
Hereditary spastic paraplegias (HSP) are characterized by spasticity and weakness of the lower limbs that result from length-dependent axonopathy of the corticospinal tracts. atlastin, the second most common HSP gene, catalyzes homotypic membrane fusion of endoplasmic reticulum (ER) tubules. How defects in neuronal Atlastin contribute to axonal degeneration has not been explained satisfactorily. Using Drosophila we demonstrate that downregulation or overexpression of Atlastin in motor neurons result in decreased crawling speed and contraction frequency in larvae, while adult flies show progressive decline in climbing ability...
August 31, 2017: Journal of Cell Science
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