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https://www.readbyqxmd.com/read/28619825/spef2-functions-in-microtubule-mediated-transport-in-elongating-spermatids
#1
Mari S Lehti, Fu-Ping Zhang, Noora Kotaja, Anu Sironen
Sperm differentiation requires specific protein transport for correct sperm tail formation and head shaping. A transient microtubular structure, the manchette, appears around the differentiating spermatid head and serves as a platform for protein transport to the growing tail. Sperm flagellar protein 2 (SPEF2) is known to be essential for sperm tail development. In this study we investigated the function of SPEF2 during spermatogenesis using a male germ cell-specific Spef2 knockout mouse model. In addition to defects in sperm tail development, we observed a duplication of the basal body and failure in the manchette migration resulting in an abnormal head shape...
June 15, 2017: Development
https://www.readbyqxmd.com/read/28576874/bloc-1-is-required-for-selective-membrane-protein-trafficking-from-endosomes-to-primary-cilia
#2
William J Monis, Victor Faundez, Gregory J Pazour
Primary cilia perceive the extracellular environment through receptors localized in the ciliary membrane, but mechanisms directing specific proteins to this domain are poorly understood. To address this question, we knocked down proteins potentially important for ciliary membrane targeting and determined how this affects the ciliary trafficking of fibrocystin, polycystin-2, and smoothened. Our analysis showed that fibrocystin and polycystin-2 are dependent on IFT20, GMAP210, and the exocyst complex, while smoothened delivery is largely independent of these components...
July 3, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28430876/ift25-an-intraflagellar-transporter-protein-dispensable-for-ciliogenesis-in-somatic-cells-is-essential-for-sperm-flagella-formation
#3
Hong Liu, Wei Li, Yong Zhang, Zhengang Zhang, Xuejun Shang, Ling Zhang, Shiyang Zhang, Yanwei Li, Andres V Somoza, Brandon Delpi, George L Gerton, James A Foster, Rex A Hess, Gregory J Pazour, Zhibing Zhang
Intraflagellar transport (IFT) is a conserved mechanism essential for the assembly and maintenance of most eukaryotic cilia and flagella. However, IFT25, a component of the IFT complex is not required for the formation of cilia in somatic tissues. In mice, the gene is highly expressed in the testis, and its expression is up-regulated during the final phase when sperm flagella are formed. To investigate the role of IFT25 in sperm flagella formation, the gene was specifically disrupted in male germ cells. All homozygous knockout mice survived to adulthood and did not show any gross abnormalities...
April 18, 2017: Biology of Reproduction
https://www.readbyqxmd.com/read/28417161/ift54-regulates-ift20-stability-but-is-not-essential-for-tubulin-transport-during-ciliogenesis
#4
Xin Zhu, Yinwen Liang, Feng Gao, Junmin Pan
Intraflagellar transport (IFT) is required for ciliogenesis by ferrying ciliary components using IFT complexes as cargo adaptors. IFT54 is a component of the IFT-B complex and is also associated with cytoplasmic microtubules (MTs). Loss of IFT54 impairs cilia assembly as well as cytoplasmic MT dynamics. The N-terminal calponin homology (CH) domain of IFT54 interacts with tubulins/MTs and has been proposed to transport tubulin during ciliogenesis, whereas the C-terminal coiled-coil (CC) domain binds IFT20. However, the precise function of these domains in vivo is not well understood...
September 2017: Cellular and Molecular Life Sciences: CMLS
https://www.readbyqxmd.com/read/28352462/fixation-methods-can-differentially-affect-ciliary-protein-immunolabeling
#5
Kiet Hua, Russell J Ferland
BACKGROUND: Primary cilia are immotile, microtubule-based organelles present on most cells. Defects in primary cilia presence/function result in a category of developmental diseases referred to as ciliopathies. As the cilia field progresses, there is a need to consider both the ciliary and extraciliary roles of cilia proteins. However, traditional fixation methods are not always suitable for examining the full range of localizations of cilia proteins. Here, we tested a variety of fixation methods with commonly used cilia markers to determine the most appropriate fixation method for different cilia proteins...
2017: Cilia
https://www.readbyqxmd.com/read/28179459/practical-method-for-targeted-disruption-of-cilia-related-genes-by-using-crispr-cas9-mediated-homology-independent-knock-in-system
#6
Yohei Katoh, Saki Michisaka, Shohei Nozaki, Teruki Funabashi, Tomoaki Hirano, Ryota Takei, Kazuhisa Nakayama
The CRISPR/Cas9 system has revolutionized genome editing in virtually all organisms. Although the CRISPR/Cas9 system enables the targeted cleavage of genomic DNA, its use for gene knock-in remains challenging because levels of homologous recombination activity vary among various cells. In contrast, the efficiency of homology-independent DNA repair is relatively high in most cell types. Therefore the use of a homology-independent repair mechanism is a possible alternative for efficient genome editing. Here we constructed a donor knock-in vector optimized for the CRISPR/Cas9 system and developed a practical system that enables efficient disruption of target genes by exploiting homology-independent repair...
April 1, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28154159/the-t-cell-ift20-interactome-reveals-new-players-in-immune-synapse-assembly
#7
Donatella Galgano, Anna Onnis, Elisa Pappalardo, Federico Galvagni, Oreste Acuto, Cosima T Baldari
Sustained signalling at the immune synapse (IS) requires the synaptic delivery of recycling endosome-associated T cell antigen receptors (TCRs). IFT20, a component of the intraflagellar transport system, controls TCR recycling to the IS as a complex with IFT57 and IFT88. Here, we used quantitative mass spectrometry to identify additional interaction partners of IFT20 in Jurkat T cells. In addition to IFT57 and IFT88, the analysis revealed new binding partners, including IFT54 (also known as TRAF3IP1), GMAP-210 (also known as TRIP11), Arp2/3 complex subunit-3 (ARPC3), COP9 signalosome subunit-1 (CSN1, also known as GPS1) and ERGIC-53 (also known as LMAN1)...
March 15, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28127051/ror2-signaling-regulates-golgi-structure-and-transport-through-ift20-for-tumor-invasiveness
#8
Michiru Nishita, Seung-Yeol Park, Tadashi Nishio, Koki Kamizaki, ZhiChao Wang, Kota Tamada, Toru Takumi, Ryuju Hashimoto, Hiroki Otani, Gregory J Pazour, Victor W Hsu, Yasuhiro Minami
Signaling through the Ror2 receptor tyrosine kinase promotes invadopodia formation for tumor invasion. Here, we identify intraflagellar transport 20 (IFT20) as a new target of this signaling in tumors that lack primary cilia, and find that IFT20 mediates the ability of Ror2 signaling to induce the invasiveness of these tumors. We also find that IFT20 regulates the nucleation of Golgi-derived microtubules by affecting the GM130-AKAP450 complex, which promotes Golgi ribbon formation in achieving polarized secretion for cell migration and invasion...
December 2017: Scientific Reports
https://www.readbyqxmd.com/read/27882921/a-mutation-in-vps15-pik3r4-causes-a-ciliopathy-and-affects-ift20-release-from-the-cis-golgi
#9
Corinne Stoetzel, Séverine Bär, Johan-Owen De Craene, Sophie Scheidecker, Christelle Etard, Johana Chicher, Jennifer R Reck, Isabelle Perrault, Véronique Geoffroy, Kirsley Chennen, Uwe Strähle, Philippe Hammann, Sylvie Friant, Hélène Dollfus
Ciliopathies are a group of diseases that affect kidney and retina among other organs. Here, we identify a missense mutation in PIK3R4 (phosphoinositide 3-kinase regulatory subunit 4, named VPS15) in a family with a ciliopathy phenotype. Besides being required for trafficking and autophagy, we show that VPS15 regulates primary cilium length in human fibroblasts, as well as ciliary processes in zebrafish. Furthermore, we demonstrate its interaction with the golgin GM130 and its localization to the Golgi. The VPS15-R998Q patient mutation impairs Golgi trafficking functions in humanized yeast cells...
November 24, 2016: Nature Communications
https://www.readbyqxmd.com/read/27682589/intraflagellar-transport-protein-ift20-is-essential-for-male-fertility-and-spermiogenesis-in-mice
#10
Zhengang Zhang, Wei Li, Yong Zhang, Ling Zhang, Maria E Teves, Hong Liu, Jerome F Strauss, Gregory J Pazour, James A Foster, Rex A Hess, Zhibing Zhang
Intraflagellar transport (IFT) is a conserved mechanism thought to be essential for the assembly and maintenance of cilia and flagella. However, little is known about its role in mammalian sperm flagella formation. To fill this gap, we disrupted the Ift20 gene in male germ cells. Homozygous mutant mice were infertile with significantly reduced sperm counts and motility. In addition, abnormally shaped elongating spermatid heads and bulbous round spermatids were found in the lumen of the seminiferous tubules...
September 28, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27678193/depletion-of-primary-cilia-from-mature-dentate-granule-cells-impairs-hippocampus-dependent-contextual-memory
#11
Soyoung Rhee, Gregory W Kirschen, Yan Gu, Shaoyu Ge
The primary cilium, a sensory organelle, regulates cell proliferation and neuronal development of dentate granule cells in the hippocampus. However, its role in the function of mature dentate granule cells remains unknown. Here we specifically depleted and disrupted ciliary proteins IFT20 and Kif3A (respectively) in mature dentate granule cells and investigated hippocampus-dependent contextual memory and long-term plasticity at mossy fiber synapses. We found that depletion of IFT20 in these cells significantly impaired context-dependent fear-related memory...
September 28, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27118846/canonical-and-noncanonical-intraflagellar-transport-regulates-craniofacial-skeletal-development
#12
Kazuo Noda, Megumi Kitami, Kohei Kitami, Masaru Kaku, Yoshihiro Komatsu
The primary cilium is a cellular organelle that coordinates signaling pathways critical for cell proliferation, differentiation, survival, and homeostasis. Intraflagellar transport (IFT) plays a pivotal role in assembling primary cilia. Disruption and/or dysfunction of IFT components can cause multiple diseases, including skeletal dysplasia. However, the mechanism by which IFT regulates skeletogenesis remains elusive. Here, we show that a neural crest-specific deletion of intraflagellar transport 20 (Ift20) in mice compromises ciliogenesis and intracellular transport of collagen, which leads to osteopenia in the facial region...
May 10, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/26715756/ift20-controls-lat-recruitment-to-the-immune-synapse-and-t-cell-activation-in-vivo
#13
Omar I Vivar, Giulia Masi, Jean-Marie Carpier, Joao G Magalhaes, Donatella Galgano, Gregory J Pazour, Sebastian Amigorena, Claire Hivroz, Cosima T Baldari
Biogenesis of the immune synapse at the interface between antigen-presenting cells and T cells assembles and organizes a large number of membrane proteins required for effective signaling through the T-cell receptor. We showed previously that the intraflagellar transport protein 20 (IFT20), a component of the intraflagellar transport system, controls polarized traffic during immune synapse assembly. To investigate the role of IFT20 in primary CD4(+) T cells in vitro and in vivo, we generated mice bearing a conditional defect of IFT20 expression in T cells...
January 12, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/26266958/chibby-functions-to-preserve-normal-ciliary-morphology-through-the-regulation-of-intraflagellar-transport-in-airway-ciliated-cells
#14
Saul S Siller, Michael C Burke, Feng-Qian Li, Ken-Ichi Takemaru
Airway cilia provide the coordinated motive force for mucociliary transport, which prevents the accumulation of mucus, debris, pollutants, and bacteria in our respiratory tracts. As airway cilia are constantly exposed to the environment and, hence, are an integral component of the pathogenesis of several congenital and chronic pulmonary disorders, it is necessary to understand the molecular mechanisms that control ciliated cell differentiation and ciliogenesis. We have previously reported that loss of the basal body protein Chibby (Cby) results in chronic upper airway infection in mice due to a significant reduction in the number of airway cilia...
2015: Cell Cycle
https://www.readbyqxmd.com/read/26157348/yes-associated-protein-yap-is-necessary-for-ciliogenesis-and-morphogenesis-during-pronephros-development-in-zebrafish-danio-rerio
#15
Liangliang He, Wenyan Xu, Ying Jing, Ming Wu, Shuwei Song, Ying Cao, Changlin Mei
The Hippo signaling pathway and its transcriptional co-activator Yap are known as essential regulators for cell proliferation and organ size. However, little is known about their roles in kidney development and ciliogenesis. We examined expression of Yap during zebrafish embryogenesis, and its transcripts were detected in pronephric duct, while Yap protein was found to be localized in the cytoplasm and apical membrane in kidney epithelium cells. By morpholino (MO) knockdown of yap expression in zebrafish, the injected larve exhibits pronephic cysts and many aspects of ciliopathy, which can be rescued by full-length yap mRNA, but not yap (S127A) mRNA...
2015: International Journal of Biological Sciences
https://www.readbyqxmd.com/read/26097753/deletion-of-ift20-in-early-stage-t-lymphocyte-differentiation-inhibits-the-development-of-collagen-induced-arthritis
#16
Xue Yuan, Lee Ann Garrett-Sinha, Debanjan Sarkar, Shuying Yang
IFT20 is the smallest member of the intraflagellar transport protein (IFT) complex B. It is involved in cilia formation. Studies of IFT20 have been confined to ciliated cells. Recently, IFT20 was found to be also expressed in non-ciliated T cells and have functions in immune synapse formation and signaling in vitro. However, how IFT20 regulates T-cell development and activation in vivo is still unknown. We deleted the IFT20 gene in early and later stages of T-cell development by crossing IFT20(flox/flox) (IFT20(f/f) ) mice with Lck-Cre and CD4-Cre transgenic mice, and investigated the role of IFT20 in T-cell maturation and in the development of T cell-mediated collagen-induced arthritis (CIA)...
2014: Bone Research
https://www.readbyqxmd.com/read/26034069/the-small-gtpase-rab8-interacts-with-vamp-3-to-regulate-the-delivery-of-recycling-t-cell-receptors-to-the-immune-synapse
#17
Francesca Finetti, Laura Patrussi, Donatella Galgano, Chiara Cassioli, Giuseppe Perinetti, Gregory J Pazour, Cosima T Baldari
IFT20, a component of the intraflagellar transport (IFT) system that controls ciliogenesis, regulates immune synapse assembly in the non-ciliated T-cell by promoting T-cell receptor (TCR) recycling. Here, we have addressed the role of Rab8 (for which there are two isoforms Rab8a and Rab8b), a small GTPase implicated in ciliogenesis, in TCR traffic to the immune synapse. We show that Rab8, which colocalizes with IFT20 in Rab11(+) endosomes, is required for TCR recycling. Interestingly, as opposed to in IFT20-deficient T-cells, TCR(+) endosomes polarized normally beneath the immune synapse membrane in the presence of dominant-negative Rab8, but were unable to undergo the final docking or fusion step...
July 15, 2015: Journal of Cell Science
https://www.readbyqxmd.com/read/26021297/the-small-gtpase-rab29-is-a-common-regulator-of-immune-synapse-assembly-and-ciliogenesis
#18
A Onnis, F Finetti, L Patrussi, M Gottardo, C Cassioli, S Spanò, C T Baldari
Accumulating evidence underscores the T-cell immune synapse (IS) as a site of intense vesicular trafficking, on which productive signaling and cell activation crucially depend. Although the T-cell antigen receptor (TCR) is known to exploit recycling to accumulate to the IS, the specific pathway that controls this process remains to be elucidated. Here we demonstrate that the small GTPase Rab29 is centrally implicated in TCR trafficking and IS assembly. Rab29 colocalized and interacted with Rab8, Rab11 and IFT20, a component of the intraflagellar transport system that regulates ciliogenesis and participates in TCR recycling in the non-ciliated T cell, as assessed by co-immunoprecipitation and immunofluorescence analysis...
October 2015: Cell Death and Differentiation
https://www.readbyqxmd.com/read/25967552/the-mal-protein-is-crucial-for-proper-membrane-condensation-at-the-ciliary-base-which-is-required-for-primary-cilium-elongation
#19
Elena Reales, Miguel Bernabé-Rubio, Javier Casares-Arias, Carles Rentero, Jaime Fernández-Barrera, Laura Rangel, Isabel Correas, Carlos Enrich, Germán Andrés, Miguel A Alonso
The base of the primary cilium contains a zone of condensed membranes whose importance is not known. Here, we have studied the involvement of MAL, a tetraspanning protein that exclusively partitions into condensed membrane fractions, in the condensation of membranes at the ciliary base and investigated the importance of these membranes in primary cilium formation. We show that MAL accumulates at the ciliary base of epithelial MDCK cells. Knockdown of MAL expression resulted in a drastic reduction in the condensation of membranes at the ciliary base, the percentage of ciliated cells and the length of the cilia, but did not affect the docking of the centrosome to the plasma membrane or produce missorting of proteins to the pericentriolar zone or to the membrane of the remaining cilia...
June 15, 2015: Journal of Cell Science
https://www.readbyqxmd.com/read/25686250/early-steps-in-primary-cilium-assembly-require-ehd1-ehd3-dependent-ciliary-vesicle-formation
#20
Quanlong Lu, Christine Insinna, Carolyn Ott, Jimmy Stauffer, Petra A Pintado, Juliati Rahajeng, Ulrich Baxa, Vijay Walia, Adrian Cuenca, Yoo-Seok Hwang, Ira O Daar, Susana Lopes, Jennifer Lippincott-Schwartz, Peter K Jackson, Steve Caplan, Christopher J Westlake
Membrane association with mother centriole (M-centriole) distal appendages is critical for ciliogenesis initiation. How the Rab GTPase Rab11-Rab8 cascade functions in early ciliary membrane assembly is unknown. Here, we show that the membrane shaping proteins EHD1 and EHD3, in association with the Rab11-Rab8 cascade, function in early ciliogenesis. EHD1 and EHD3 localize to preciliary membranes and the ciliary pocket. EHD-dependent membrane tubulation is essential for ciliary vesicle formation from smaller distal appendage vesicles (DAVs)...
March 2015: Nature Cell Biology
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