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Dhiman Ghosh, Ilya V Ulasov, LiPing Chen, Lualhati E Harkins, Karolina Wallenborg, Parvinder Hothi, Steven Rostad, Leroy Hood, Charles S Cobbs
Glioblastoma multiforme (GBM) is the most common and lethal adult brain tumor. Resistance to standard radiation and chemotherapy is thought to involve survival of GBM cancer stem cells (CSCs). To date, no single marker for identifying GBM CSCs has been able to capture the diversity of CSC populations, justifying the needs for additional CSC markers for better characterization. Employing targeted mass spectrometry, here we present five cell-surface markers HMOX1, SLC16A1, CADM1, SCAMP3, and CLCC1 which were found to be elevated in CSCs relative to healthy neural stem cells (NSCs)...
September 2016: Stem Cells
Thomas Falguières, David Castle, Jean Gruenberg
The biogenesis of multivesicular endosomes and the sorting of activated signaling receptors into multivesicular endosomes depend on soluble protein complexes (ESCRT complexes), which transiently interact with the receptor cargo and the endosomal membrane. Previously, it was shown that the transmembrane protein secretory carrier membrane protein (SCAMP) 3, which is present on endosomes, interacts with ESCRT components. Here, we report that SCAMP3 plays a role in the biogenesis of multivesicular endosomes. We find that SCAMP3 plays a role in EGF receptor sorting into multivesicular endosomes and in the formation of intralumenal vesicles within these endosomes in vitro and thus also controls EGF receptor targeting to lysosomes...
January 2012: Traffic
Natalia A Kaniuk, Veronica Canadien, Richard D Bagshaw, Malina Bakowski, Virginie Braun, Marija Landekic, Shuvadeep Mitra, Ju Huang, Won Do Heo, Tobias Meyer, Laurence Pelletier, Helene Andrews-Polymenis, Michael McClelland, Tony Pawson, Sergio Grinstein, John H Brumell
The facultative intracellular pathogen Salmonella enterica serovar Typhimurium establishes a replicative niche, the Salmonella-containing vacuole (SCV), in host cells. Here we demonstrate that these bacteria exploit the function of Arl8B, an Arf family GTPase, during infection. Following infection, Arl8B localized to SCVs and to tubulated endosomes that extended along microtubules in the host cell cytoplasm. Arl8B(+) tubules partially colocalized with LAMP1 and SCAMP3. Formation of LAMP1(+) tubules (the Salmonella-induced filaments phenotype; SIFs) required Arl8B expression...
November 2011: Cellular Microbiology
Guoan Zhang, Katrin Deinhardt, Moses V Chao, Thomas A Neubert
Conventional stable isotope labeling with amino acids in cell culture (SILAC) requires extensive metabolic labeling of proteins and therefore is difficult to apply to cells that do not divide or are unstable in SILAC culture. Using two different sets of heavy amino acids for labeling allows for straightforward SILAC quantitation using partially labeled cells because the two cell populations are always equally labeled. Here we report the application of this labeling strategy to primary cultured neurons. We demonstrated that protein quantitation was not compromised by incomplete labeling of the neuronal proteins...
May 6, 2011: Journal of Proteome Research
Nina Schroeder, Luís Jaime Mota, Stéphane Méresse
Salmonella virulence relies on its capacity to replicate inside various cell types in a membrane-bound compartment, the Salmonella-containing vacuole (SCV). A unique feature of Salmonella-infected cells is the presence of tubular structures originating from and connected to the SCV, which often extend throughout the cell cytoplasm. These tubules include the well-studied Salmonella-induced filaments (SIFs), enriched in lysosomal membrane proteins. However, recent studies revealed that the Salmonella-induced tubular network is more extensive than previously thought and includes three types of tubules distinct from SIFs: sorting nexin tubules, Salmonella-induced secretory carrier membrane protein 3 (SCAMP3) tubules and lysosome-associated membrane protein 1 (LAMP1)-negative tubules...
June 2011: Trends in Microbiology
Luís Jaime Mota, Amy E Ramsden, Mei Liu, J David Castle, David W Holden
Salmonella enterica are facultative intracellular bacterial pathogens that proliferate within host cells in a membrane-bounded compartment, the Salmonella-containing vacuole (SCV). Intracellular replication of Salmonella is mediated by bacterial effectors translocated on to the cytoplasmic face of the SCV membrane by a type III secretion system. Some of these effectors manipulate the host endocytic pathway, resulting in the formation in epithelial cells of tubules enriched in late endosomal markers, known as Salmonella-induced filaments (SIFs)...
August 2009: Cellular Microbiology
Quyen L Aoh, Anna M Castle, Charles H Hubbard, Osamu Katsumata, J David Castle
The epidermal growth factor receptor (EGFR) is targeted for lysosomal degradation by ubiquitin-mediated interactions with the ESCRTs (endosomal-sorting complexes required for transport) in multivesicular bodies (MVBs). We show that secretory carrier membrane protein, SCAMP3, localizes in part to early endosomes and negatively regulates EGFR degradation through processes that involve its ubiquitylation and interactions with ESCRTs. SCAMP3 is multimonoubiquitylated and is able to associate with Nedd4 HECT ubiquitin ligases and the ESCRT-I subunit Tsg101 via its PY and PSAP motifs, respectively...
March 2009: Molecular Biology of the Cell
Hua Wang, Nicholas P Hays, Swapan K Das, Rebekah L Craig, Winston S Chu, Neeraj Sharma, Steven C Elbein
OBJECTIVE: Linkage to type 2 diabetes (T2D) is well replicated on chromosome 1q21-q23. Within this region, T2D was associated with common single nucleotide polymorphisms that marked an extended linkage disequilibrium block, including the liver pyruvate kinase gene (PKLR), in several European-derived populations. In this study we sought to determine the molecular basis for the association and the phenotypic consequences of the risk haplotype. RESEARCH DESIGN AND METHODS: Genes surrounding PKLR were resequenced in European-American and African-American cases and controls, and association with T2D was tested...
April 2009: Journal of Clinical Endocrinology and Metabolism
Galih Kunarso, Kee-Yew Wong, Lawrence W Stanton, Leonard Lipovich
BACKGROUND: Transcriptional control of embryonic stem (ES) cell pluripotency has been a subject of intense study. Transcriptional regulators including Oct4 (Oct3/4 index), Sox2 and Nanog are fundamental for maintaining the undifferentiated state. However, the ES cell transcriptome is not limited to their targets, and exhibits considerable complexity when assayed with microarray, MPSS, cDNA/EST sequencing, and SAGE technologies. To identify novel genes associated with pluripotency, we globally searched for ES transcripts not corresponding to known genes, validated their sequences, determined their expression profiles, and employed RNAi to test their function...
2008: BMC Genomics
Britta Skawran, Doris Steinemann, Anja Weigmann, Peer Flemming, Thomas Becker, Jakobus Flik, Hans Kreipe, Brigitte Schlegelberger, Ludwig Wilkens
Cytogenetics of hepatocellular carcinoma and adenoma have revealed gains of chromosome 1q as a significant differentiating factor. However, no studies are available comparing these amplification events with gene expression. Therefore, gene expression profiling was performed on tumours cytogenetically well characterized by array-based comparative genomic hybridisation. For this approach analysis was carried out on 24 hepatocellular carcinoma and 8 hepatocellular adenoma cytogenetically characterised by array-based comparative genomic hybridisation...
May 2008: Modern Pathology: An Official Journal of the United States and Canadian Academy of Pathology, Inc
Matthias T F Wolf, Bettina E Mucha, Hans C Hennies, Massimo Attanasio, Franziska Panther, Isabella Zalewski, Stephanie M Karle, Edgar A Otto, C Constantinou Deltas, Arno Fuchshuber, Friedhelm Hildebrandt
Medullary cystic kidney disease type 1 (MCKD1) is an autosomal dominant, tubulo-interstitial nephropathy that causes renal salt wasting and end-stage renal failure in the fourth to seventh decade of life. MCKD1 was localized to chromosome 1q21. We demonstrated haplotype sharing and confirmed the telomeric border by a recombination of D1S2624 in a Belgian kindred. Since the causative gene has been elusive, high resolution haplotype analysis was performed in 16 kindreds. Clinical data and blood samples of 257 individuals (including 75 affected individuals) from 26 different kindreds were collected...
July 2006: Human Genetics
T T Wu, J D Castle
Secretory carrier membrane proteins (SCAMPs) are ubiquitously expressed proteins of post-Golgi vesicles. In the presence of the tyrosine phosphatase inhibitor vanadate, or after overexpression in Chinese hamster ovary (CHO) cells, SCAMP1 and SCAMP3 are phosphorylated selectively on tyrosine residue(s). Phosphorylation is reversible after vanadate washout in situ or when isolated SCAMP3 is incubated with the recombinant tyrosine phosphatase PTP1B. Vanadate also causes the partial accumulation of SCAMP3, but not SCAMP1, in "patches" at or near the cell surface...
July 1998: Molecular Biology of the Cell
D R Singleton, T T Wu, J D Castle
The primary structures of three human forms of secretory carrier membrane proteins (SCAMPs) have been deduced from full-length clones isolated from a HeLa cell cDNA library and confirmed by a combination of comparison to expressed sequence tags, microsequencing of purified protein, and in vitro transcription and translation. The structures indicated that SCAMPs are highly related products of distinct genes, and that the sequence identity of an individual SCAMP between different mammalian species is almost complete...
September 1997: Journal of Cell Science
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