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https://www.readbyqxmd.com/read/29218523/identification-of-insulin-activated-rab-proteins-in-adipose-cells-using-bio-atb-gtp-photolabeling-technique
#1
Francoise Koumanov, Geoffrey D Holman
We have recently developed a photolabeling method to identify GTP-loaded Rab proteins. The new biotinylated GTP analogue (Bio-ATB-GTP) binds to GTP-binding proteins and after a UV irradiation a covalent bond is formed between the protein and the photoreactive diazirine group on the photolabel. The tagged protein can then be isolated and detected using the classic biotin-streptavidin interaction. In this chapter, we describe the Bio-ATB-GTP photolabel and discuss the advantages of using this photolabeling approach to detect GTP-loaded Rab proteins compared to other existing methodologies...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29215055/impairing-the-function-of-mlck-myosin-va-or-myosin-vb-disrupts-rhinovirus-b14-replication
#2
Antonio Real-Hohn, D William Provance, Rafael Braga Gonçalves, Caio Bidueira Denani, Andréa Cheble de Oliveira, Verônica P Salerno, Andre Marco Oliveira Gomes
Together, the three human rhinovirus (RV) species are the most frequent cause of the common cold. Because of their high similarity with other viral species of the genus Enterovirus, within the large family Picornaviridae, studies on RV infectious activities often offer a less pathogenic model for more aggressive enteroviruses, e.g. poliovirus or EV71. Picornaviruses enter via receptor mediated endocytosis and replicate in the cytosol. Most of them depend on functional F-actin, Rab proteins, and probably motor proteins...
December 7, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29212907/role-of-the-ehd-family-of-endocytic-recycling-regulators-for-tcr-recycling-and-t-cell-function
#3
Fany M Iseka, Benjamin T Goetz, Insha Mushtaq, Wei An, Luke R Cypher, Timothy A Bielecki, Eric C Tom, Priyanka Arya, Sohinee Bhattacharyya, Matthew D Storck, Craig L Semerad, James E Talmadge, R Lee Mosley, Vimla Band, Hamid Band
T cells use the endocytic pathway for key cell biological functions, including receptor turnover and maintenance of the immunological synapse. Some of the established players include the Rab GTPases, the SNARE complex proteins, and others, which function together with EPS-15 homology domain-containing (EHD) proteins in non-T cell systems. To date, the role of the EHD protein family in T cell function remains unexplored. We generated conditional EHD1/3/4 knockout mice using CD4-Cre and crossed these with mice bearing a myelin oligodendrocyte glycoprotein-specific TCR transgene...
December 6, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/29212874/the-na-k-h-exchanger-nhx1-controls-multivesicular-body-vacuolar-lysosome-fusion
#4
Mahmoud Abdul Karim, Christopher Leonard Brett
Loss-of-function mutations in human endosomal Na+(K+)/H+ Exchangers (NHEs) NHE6 and NHE9 are implicated in neurological disorders including Christianson Syndrome, autism and attention deficit and hyperactivity disorder (ADHD). These mutations disrupt retention of surface receptors within neurons and glial cells by affecting their delivery to lysosomes for degradation. However, the molecular basis of how these endosomal NHEs control endocytic trafficking is unclear. Using Saccharomyces cerevisiae as a model, we conducted cell-free organelle fusion assays to show that transport activity of the orthologous endosomal NHE Nhx1 is important for multivesicular body (MVB)-vacuolar lysosome fusion, the last step of endocytosis required for surface protein degradation...
December 6, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/29212815/rab29-activation-of-the-parkinson-s-disease-associated-lrrk2-kinase
#5
Elena Purlyte, Herschel S Dhekne, Adil R Sarhan, Rachel Gomez, Pawel Lis, Melanie Wightman, Terina N Martinez, Francesca Tonelli, Suzanne R Pfeffer, Dario R Alessi
Parkinson's disease predisposing LRRK2 kinase phosphorylates a group of Rab GTPase proteins including Rab29, within the effector-binding switch II motif. Previous work indicated that Rab29, located within the PARK16 locus mutated in Parkinson's patients, operates in a common pathway with LRRK2. Here, we show that Rab29 recruits LRRK2 to the trans-Golgi network and greatly stimulates its kinase activity. Pathogenic LRRK2 R1441G/C and Y1699C mutants that promote GTP binding are more readily recruited to the Golgi and activated by Rab29 than wild-type LRRK2...
December 6, 2017: EMBO Journal
https://www.readbyqxmd.com/read/29212243/effects-of-silencing-rab27a-gene-on-biological-characteristics-and-chemosensitivity-of-non-small-cell-lung-cancer
#6
Xia Li, Haiying Wang, Qinggan Ni, Zhiyuan Tang, Jun Ni, Liqin Xu, Hua Huang, Songshi Ni, Jian Feng
Rab27a, a member of the Rab protein family, can regulate the tumor microenvironment and promote the development of the tumor. Elevated expression of Rab27a is closely connected with many human cancers containing non-small cell lung cancer (NSCLC). But the role of Rab27a in non-small cell lung cancer and its possible mechanism is particularly unclear. In this research, we explored the effect of silencing Rab27a in vitro and in vivo, furnishing evidence that Rab27a could be a potential therapeutic target in NSCLC...
November 7, 2017: Oncotarget
https://www.readbyqxmd.com/read/29208657/assembly-of-intermediates-for-rapid-membrane-fusion
#7
Max Harner, William Wickner
Membrane fusion is essential for intracellular protein sorting, cell growth, hormone secretion, and neurotransmission. Rapid membrane fusion requires tethering and Sec1-Munc18 (SM) function to catalyze R-, Qa-, Qb-, and Qc-SNARE complex assembly in trans, as well as SNARE engagement by the SNARE-binding chaperone Sec17/αSNAP. The hexameric vacuolar HOPS complex in the yeast Saccharomyces cerevisiae tethers membranes through its affinities for the membrane Rab GTPase Ypt7. HOPS also has specific affinities for the vacuolar SNAREs and catalyzes SNARE complex assembly, but the order of their assembly into a 4-SNARE complex is unclear...
December 5, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29204879/reconstitution-of-membrane-tethering-mediated-by-rab-family-small-gtpases
#8
REVIEW
Joji Mima
Membrane tethering is one of the most critical steps to determine the spatiotemporal specificity of membrane trafficking, which is the process to selectively transport proteins, lipids, and other biological molecules to the appropriate locations in eukaryotic cells, such as subcellular organelles, the plasma membrane, and the extracellular space. Based on genetic, cell biological, biochemical, and structural studies, Rab-family small GTPases and a number of Rab-interacting proteins (termed Rab effectors), including coiled-coil tethering proteins and multisubunit tethering complexes, have been proposed to be key protein components for membrane tethering...
December 4, 2017: Biophysical Reviews
https://www.readbyqxmd.com/read/29204810/high-risk-palliative-care-patients-knowledge-and-attitudes-about-hereditary-cancer-testing-and-dna-banking
#9
John M Quillin, Oluwabunmi Emidio, Brittany Ma, Lauryn Bailey, Thomas J Smith, In Guk Kang, Brandon J Yu, Oluwafemi Patrick Owodunni, Mohammed Abusamaan, Rab Razzak, Joann N Bodurtha
Even at the end of life, testing cancer patients for inherited susceptibility may provide life-saving information to their relatives. Prior research suggests palliative care inpatients have suboptimal understanding of genetic importance, and testing may be underutilized in this clinical setting. These conclusions are based on limited research. This study aimed to estimate genetic testing prevalence among high-risk palliative care patients in a National Cancer Institute-designated comprehensive cancer center...
December 4, 2017: Journal of Genetic Counseling
https://www.readbyqxmd.com/read/29201946/cell-free-fluorescent-intra-golgi-retrograde-vesicle-trafficking-assay
#10
Nathanael P Cottam, Daniel Ungar
Intra-Golgi retrograde vesicle transport is used to traffic and sort resident Golgi enzymes to their appropriate cisternal locations. An assay was established to investigate the molecular details of vesicle targeting in a cell-free system. Stable cell lines were generated in which the trans-Golgi enzyme galactosyltransferase (GalT) was tagged with either CFP or YFP. Given that GalT is recycled to the cisterna where it is located at steady state, GalT-containing vesicles target GalT-containing cisternal membranes...
November 20, 2017: Bio-protocol
https://www.readbyqxmd.com/read/29196555/ral-gtpases-biology-and-potential-as-therapeutic-targets-in-cancer
#11
REVIEW
Chao Yan, Dan Theodorescu
More than a hundred proteins comprise the RAS superfamily of small GTPases. This family can be divided into RAS, RHO, RAB, RAN, ARF, and RAD subfamilies, with each shown to play distinct roles in human cells in both health and disease. The RAS subfamily has a well-established role in human cancer with the three genes, HRAS, KRAS, and NRAS being the commonly mutated in tumors. These RAS mutations, most often functionally activating, are especially common in pancreatic, lung, and colorectal cancers. Efforts to inhibit RAS and related GTPases have produced inhibitors targeting the downstream effectors of RAS signaling, including inhibitors of the RAF-mitogen-activated protein kinase/extracellular signal-related kinase (ERK)-ERK kinase pathway and the phosphoinositide-3-kinase-AKT-mTOR kinase pathway...
January 2018: Pharmacological Reviews
https://www.readbyqxmd.com/read/29193252/immune-function-of-a-rab-related-protein-by-modulating-the-jak-stat-signaling-pathway-in-the-silkworm-bombyx-mori
#12
Chen Chen, Salah Eldein, Xiaosan Zhou, Yu Sun, Jin Gao, Yuxuan Sun, Chaoliang Liu, Lei Wang
The Rab-family GTPases mainly regulate intracellular vesicle transport, and play important roles in the innate immune response in invertebrates. However, the function and signal transduction of Rab proteins in immune reactions remain unclear in silkworms. In this study, we analyzed a Rab-related protein of silkworm Bombyx mori (BmRABRP) by raising antibodies against its bacterially expressed recombinant form. Tissue distribution analysis showed that BmRABRP mRNA and protein were high expressed in the Malpighian tubule and fat body, respectively...
November 30, 2017: Archives of Insect Biochemistry and Physiology
https://www.readbyqxmd.com/read/29188508/retinal-gene-therapy-for-choroideremia-in-vitro-testing-for-gene-augmentation-using-an-adeno-associated-viral-aav-vector
#13
Maria I Patrício, Robert E MacLaren
As gene therapy of choroideremia is becoming a clinical reality, there is a need for reliable and sensitive assays to determine the expression of exogenously delivered Rab Escort Protein-1 (REP1), in particular to test new gene therapy vectors and as a quality control screen for clinical vector stocks. Here we describe an in vitro protocol to test transgene expression following AAV2/2-REP1 transduction of a human cell line. Gene augmentation can be confirmed by western blot and quantification of the fold-increase of human REP1 levels over untransduced controls...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29184002/a-guanine-nucleotide-exchange-factor-gef-limits-rab-gtpase-driven-membrane-fusion
#14
Lars Langemeyer, Angela Perz, Daniel Kuemmel, Christian Ungermann
The identity of organelles in the endomembrane system of any eukaryotic cell critically depends on the correctly localized Rab GTPase, which binds effectors and thus promotes membrane remodeling or fusion. However, it is still unresolved which factors are required and therefore define the localization of the correct fusion machinery. Using SNARE-decorated proteoliposomes that cannot fuse on their own, we now demonstrate that full fusion activity can be achieved by just four soluble factors: a soluble SNARE (V am7), a guanine nucleotide exchange factor (GEF, Mon1-Ccz1), a Rab- GDP dissociation inhibitor (GDI) complex (prenylated Ypt7:GDI), and a Rab effector complex (HOPS)...
November 28, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29178045/considering-choline-as-methionine-precursor-lipoproteins-transporter-hepatic-promoter-and-antioxidant-agent-in-dairy-cows
#15
REVIEW
Imtiaz Hussain Raja Abbasi, Farzana Abbasi, Rab N Soomro, Mohamed E Abd El-Hack, Mervat A Abdel-Latif, Wen Li, Ren Hao, Feifei Sun, Bello M Bodinga, Khawar Hayat, Junhu Yao, Yangchun Cao
During the transition period, fatty liver syndrome may be caused in cows undergo negative energy balance, ketosis or hypocalcemia, retained placenta or mastitis problems. During the transition stage, movement of non-esterified fatty acids (NEFA) increases into blood which declines the hepatic metabolism or reproduction and consequently, lactation performance of dairy cows deteriorates. Most of studies documented that, choline is an essential nutrient which plays a key role to decrease fatty liver, NEFA proportion, improve synthesis of phosphatidylcholine, maintain lactation or physiological function and work as anti-oxidant in the transition period of dairy cows...
November 25, 2017: AMB Express
https://www.readbyqxmd.com/read/29177506/lrrk2-phosphorylates-membrane-bound-rabs-and-is-activated-by-gtp-bound-rab7l1-to-promote-recruitment-to-the-trans-golgi-network
#16
Zhiyong Liu, Nicole Bryant, Ravindran Kumaran, Alexandra Beilina, Asa Abeliovich, Mark R Cookson, Andrew B West
Human genetic studies implicate LRRK2 and Rab7L1 in susceptibility to Parkinson disease (PD). These two genes function in the same pathway, as knockout of Rab7L1 results in phenotypes similar to LRRK2 knockout, and studies in cells and model organisms demonstrate LRRK2 and Rab7L1 interact in the endolysosomal system. Recently, a subset of Rab proteins have been identified as LRRK2 kinase substrates. Herein, we find that Rab8, Rab10, and Rab7L1 must be membrane and GTP-bound for LRRK2 phosphorylation. LRRK2 mutations that cause PD including R1441C, Y1699C, and G2019S all increase LRRK2 phosphorylation of Rab7L1 four-fold over wild-type LRRK2 in cells, resulting in the phosphorylation of nearly one-third the available Rab7L1 protein in cells...
November 21, 2017: Human Molecular Genetics
https://www.readbyqxmd.com/read/29167152/rab27b-requirement-for-stretch-induced-exocytosis-in-bladder-umbrella-cells
#17
Luciana I Gallo, Marianela G Dalghi, Dennis R Clayton, Wily G Ruiz, Puneet Khandelwal, Gerard Apodaca
Umbrella cells, which must maintain a tight barrier, modulate their apical surface area during bladder filling by exocytosis of an abundant, subapical pool of discoidal- and/or fusiform-shaped vesicles (DFVs). Despite the importance of this trafficking event for bladder function, the pathways the promote DFV exocytosis remain to be identified. We previously showed that DFV exocytosis depends in part on a RAB11A-RAB8A-MYO5B network, but RAB27B is also reported to be associated with DFVs, and knockout mice lacking RAB27B have fewer DFVs...
November 22, 2017: American Journal of Physiology. Cell Physiology
https://www.readbyqxmd.com/read/29166671/the-15n-leucine-single-injection-method-allows-for-determining-endogenous-losses-and-true-digestibility-of-amino-acids-in-cecectomized-roosters
#18
Rujiu Hu, Jing Li, Rab Nawaz Soomro, Fei Wang, Yan Feng, Xiaojun Yang, Junhu Yao
This study was conducted to assess the influence of dietary protein content in poultry when using the 15N-leucine single-injection method to determine endogenous amino acid losses (EAALs) in poultry. Forty-eight cecectomized roosters (2.39 ± 0.23 kg) were randomly allocated to eight dietary treatments containing protein levels of 0, 3%, 6%, 9%, 12%, 15%, 18% and 21%. Each bird was precisely fed an experimental diet of 25 g/kg of body weight. After feeding, all roosters were subcutaneously injected with a 15N-leucine solution at a dose of 20 mg/kg of body weight...
2017: PloS One
https://www.readbyqxmd.com/read/29165603/all-roads-lead-to-the-vacuole-autophagic-transport-as-part-of-the-endomembrane-trafficking-network-in-plants
#19
Kamila Kalinowska, Erika Isono
Plants regulate their development and response to the changing environment by sensing and interpreting environmental signals. Intracellular trafficking pathways including endocytic-, vacuolar-, and autophagic trafficking are important for the various aspects of responses in plants. Studies in the last decade have shown that the autophagic transport pathway uses common key components of endomembrane trafficking as well as specific regulators. A number of factors previously described for their function in endosomal trafficking have been discovered to be involved in the regulation of autophagy in plants...
November 19, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/29161562/membrane-traffic-trans-golgi-tethers-leave-a%C3%A2-surprisingly-small-gap
#20
Francis A Barr
Activation and inactivation of Rab GTPases by GEFs and GAPs promotes or terminates vesicle tethering to organelles, respectively. This simple model is challenged by new evidence revealing that a catalytically inactive Rab GAP promotes rather than terminates vesicle tethering at the trans-Golgi.
November 20, 2017: Current Biology: CB
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