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A-549

RRID:CVCL_0023

Organism

Homo sapiens

Comments

Part of: Cancer Cell Line Encyclopedia (CCLE) project. Part of: ENCODE project common cell types; tier 2. Part of: JFCR39 cancer cell line panel. Part of: KuDOS 95 cell line panel. Part of: MD Anderson Cell Lines Project. Part of: Naval Biosciences Laboratory (NBL) collection (transferred to ATCC in 1982). Part of: NCI60 cancer cell line panel. Doubling time: 18 hours (in RPMI 1640+10% FBS), 37 hours (in ACL-3), 36 hours (in ACL-3+BSA) (PubMed=3940644); 22 hours (PubMed=25984343); 27 hours (from cell counting), 27 hours (from absorbance) (DOI=10.5897/IJBMBR2013.0154); 22.9 hours (NCI-DTP); ~40 hours (DSMZ). Microsatellite instability: Stable (MSS) (PubMed=12661003; Sanger). Sequence variation: Homozygous for KRAS p.Gly12Ser (PubMed=17088437). Sequence variation: Homozygous for STK11 p.Gln37Ter (PubMed=17088437). Omics: Acetylation analysis by proteomics. Omics: Deep antibody staining analysis. Omics: Deep exome analysis. Omics: Deep phosphoproteome analysis. Omics: Deep membrane proteome analysis. Omics: Deep proteome analysis. Omics: Deep RNAseq analysis. Omics: DNA methylation analysis. Omics: Fluorescence phenotype profiling. Omics: H3K4me3 ChIP-seq epigenome analysis. Omics: H3K9ac ChIP-seq epigenome analysis. Omics: lncRNA expression profiling. Omics: Metabolome analysis. Omics: Protein expression by reverse-phase protein arrays. Omics: Proteome analysis by 2D-DE/MS. Omics: shRNA library screening. Omics: SNP array analysis. Omics: Transcriptome analysis. Omics: Virome analysis using proteomics. Misspelling: 'A594' in PubMed=18227028. Misspelling: 'A59' in PubMed=16354588. Discontinued: ATCC; CRL-7909.

Proper Citation

ATCC Cat# CCL-185, RRID:CVCL_0023

Category

Cancer cell line

Sex

Male

Synonyms

A 549, A549, NCI-A549, A549/ATCC, A549 ATCC, A549ATCC, hA549

Vendor

ATCC

Cat Num

CCL-185

Cross References

AS CVCL_H249 BTO; BTO:0000018 CLO; CLO_0001601 CLO; CLO_0050025 EFO; EFO_0001086 CLDB; cl202 CLDB; cl203 CLDB; cl204 CLDB; cl205 CLDB; cl206 CLDB; cl207 CLDB; cl5100 CLDB; cl7131 AddexBio; C0016002/23 ATCC; CCL-185 ATCC; CRM-CCL-185 ATCC; CRL-7909 BCRC; 60074 BCRJ; 0033 BioSample; SAMN01821615 BioSample; SAMN03472926 CCLE; A549_LUNG CCRID; 3111C0001CCC000002 CCRID; 3111C0002000000008 CCRID; 3131C0001000700005 CCRID; 3131C0001000700150 CCRID; 3142C0001000000062 CCRID; 3153C0001000000062 CGH-DB; 202-1 CGH-DB; 9164-4 ChEMBL-Cells; CHEMBL3307651 ChEMBL-Targets; CHEMBL392 CLS; 300114/p686_A-549 Cosmic; 713864 Cosmic; 716170 Cosmic; 719675 Cosmic; 735605 Cosmic; 755468 Cosmic; 801571 Cosmic; 844592 Cosmic; 844826 Cosmic; 875854 Cosmic; 876152 Cosmic; 877443 Cosmic; 889090 Cosmic; 905949 Cosmic; 910552 Cosmic; 910560 Cosmic; 914947 Cosmic; 917386 Cosmic; 917972 Cosmic; 929148 Cosmic; 929962 Cosmic; 932915 Cosmic; 961833 Cosmic; 974287 Cosmic; 1004698 Cosmic; 1017830 Cosmic; 1028950 Cosmic; 1047097 Cosmic; 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GSM651560 GEO; GSM651561 GEO; GSM729842 GEO; GSM729843 GEO; GSM729844 GEO; GSM729845 GEO; GSM729846 GEO; GSM729847 GEO; GSM729848 GEO; GSM729849 GEO; GSM750779 GEO; GSM750780 GEO; GSM750788 GEO; GSM750789 GEO; GSM750791 GEO; GSM784244 GEO; GSM794260 GEO; GSM799336 GEO; GSM799399 GEO; GSM816649 GEO; GSM827464 GEO; GSM846285 GEO; GSM843439 GEO; GSM886858 GEO; GSM887923 GEO; GSM923418 GEO; GSM923420 GEO; GSM923425 GEO; GSM923432 GEO; GSM923433 GEO; GSM923435 GEO; GSM945243 GEO; GSM945244 GEO; GSM1086289 GEO; GSM1086290 GEO; GSM1153406 GEO; GSM1181254 GEO; GSM1181311 GEO; GSM1181344 GEO; GSM1181353 GEO; GSM1181354 GEO; GSM1181355 GEO; GSM1181356 GEO; GSM1181364 GEO; GSM1374385 GEO; GSM1374386 GEO; GSM1374387 GEO; GSM1557133 GEO; GSM1669589 GEO; GSM2124637 ICLC; HTL03001 IFO; IFO50153 IGRhCellID; A549GEO IZSLER; BS TCL 101 JCRB; JCRB0076 KCB; KCB 200434YJ KCLB; 10185 LINCS_HMS; 50084 LINCS_LDP; LCL-1601 Lonza; 675 NCBI_Iran; C137 NCI-DTP; A549 PRIDE; PXD000418 PRIDE; PXD001548 PRIDE; PXD002224 PRIDE; PXD002395 PRIDE; PXD003627 RCB; RCB0098 RCB; RCB3677 SKY/M-FISH/CGH; 2809 TKG; TKG 0184 TOKU-E; 3615 Wikidata; Q4649475

Legionella pneumophila Modulates Mitochondrial Dynamics to Trigger Metabolic Repurposing of Infected Macrophages.

  • Escoll P
  • Cell Host Microbe
  • 2017 Aug 29

Literature context: inoma cells ATCC RRID:CVCL_0023 Biological Samples


Abstract:

The intracellular bacteria Legionella pneumophila encodes a type IV secretion system (T4SS) that injects effector proteins into macrophages in order to establish and replicate within the Legionella-containing vacuole (LCV). Once generated, the LCV interacts with mitochondria through unclear mechanisms. We show that Legionella uses both T4SS-independent and T4SS-dependent mechanisms to respectively interact with mitochondria and induce mitochondrial fragmentation that ultimately alters mitochondrial metabolism. The T4SS effector MitF, a Ran GTPase activator, is required for fission of the mitochondrial network. These effects of MitF occur through accumulation of mitochondrial DNM1L, a GTPase critical for fission. Furthermore mitochondrial respiration is abruptly halted in a T4SS-dependent manner, while T4SS-independent upregulation of cellular glycolysis remains elevated. Collectively, these alterations in mitochondrial dynamics promote a Warburg-like phenotype in macrophages that favors bacterial replication. Hence the rewiring of cellular bioenergetics to create a replication permissive niche in host cells is a virulence strategy of L. pneumophila.

Environmental cystine drives glutamine anaplerosis and sensitizes cancer cells to glutaminase inhibition.

  • Muir A
  • Elife
  • 2017 Aug 15

Literature context: ) (A549: ATCC Cat# CRM-CCL-185, RRID:CVCL_0023; HCT116: ATCC Cat# CCL-247, RRI


Abstract:

Many mammalian cancer cell lines depend on glutamine as a major tri-carboxylic acid (TCA) cycle anaplerotic substrate to support proliferation. However, some cell lines that depend on glutamine anaplerosis in culture rely less on glutamine catabolism to proliferate in vivo. We sought to understand the environmental differences that cause differential dependence on glutamine for anaplerosis. We find that cells cultured in adult bovine serum, which better reflects nutrients available to cells in vivo, exhibit decreased glutamine catabolism and reduced reliance on glutamine anaplerosis compared to cells cultured in standard tissue culture conditions. We find that levels of a single nutrient, cystine, accounts for the differential dependence on glutamine in these different environmental contexts. Further, we show that cystine levels dictate glutamine dependence via the cystine/glutamate antiporter xCT/SLC7A11. Thus, xCT/SLC7A11 expression, in conjunction with environmental cystine, is necessary and sufficient to increase glutamine catabolism, defining important determinants of glutamine anaplerosis and glutaminase dependence in cancer.

mTORC2 Regulates Amino Acid Metabolism in Cancer by Phosphorylation of the Cystine-Glutamate Antiporter xCT.

  • Gu Y
  • Mol. Cell
  • 2017 Jul 6

Literature context: B-126Human: MDA-MB-231ATCCHTB-26Human: A549ATCCCCL-185Human: HEK293TATCCCRL-3216Mouse:


Abstract:

Mutations in cancer reprogram amino acid metabolism to drive tumor growth, but the molecular mechanisms are not well understood. Using an unbiased proteomic screen, we identified mTORC2 as a critical regulator of amino acid metabolism in cancer via phosphorylation of the cystine-glutamate antiporter xCT. mTORC2 phosphorylates serine 26 at the cytosolic N terminus of xCT, inhibiting its activity. Genetic inhibition of mTORC2, or pharmacologic inhibition of the mammalian target of rapamycin (mTOR) kinase, promotes glutamate secretion, cystine uptake, and incorporation into glutathione, linking growth factor receptor signaling with amino acid uptake and utilization. These results identify an unanticipated mechanism regulating amino acid metabolism in cancer, enabling tumor cells to adapt to changing environmental conditions.

Funding information:
  • NCI NIH HHS - F31 CA186668()
  • NIGMS NIH HHS - R01 GM116897()
  • NINDS NIH HHS - R01 NS073831()

A Splice Variant of Centrosomin Converts Mitochondria to Microtubule-Organizing Centers.

  • Chen JV
  • Curr. Biol.
  • 2017 Jul 10

Literature context: 8Human: U2OS cellsATCCCat#HTB-96Human: A549 cellsATCCCat#CCL-185Cercopithecus aethiops: Vero Cel


Abstract:

Non-centrosomal microtubule organizing centers (MTOCs) direct microtubule (MT) organization to exert diverse cell-type-specific functions. In Drosophila spermatids, the giant mitochondria provide structural platforms for MT reorganization to support elongation of the extremely long sperm. However, the molecular basis for this mitochondrial MTOC and other non-centrosomal MTOCs has not been discerned. Here we report that Drosophila centrosomin (cnn) expresses two major protein variants: the centrosomal form (CnnC) and a non-centrosomal form in testes (CnnT). CnnC is established as essential for functional centrosomes, the major MTOCs in animal cells. We show that CnnT is expressed exclusively in testes by alternative splicing and localizes to giant mitochondria in spermatids. In cell culture, CnnT targets to the mitochondrial surface, recruits the MT nucleator γ-tubulin ring complex (γ-TuRC), and is sufficient to convert mitochondria to MTOCs independent of core pericentriolar proteins that regulate MT assembly at centrosomes. We mapped two separate domains in CnnT: one that is necessary and sufficient to target it to mitochondria and another that is necessary and sufficient to recruit γ-TuRCs and nucleate MTs. In elongating spermatids, CnnT forms speckles on the giant mitochondria that are required to recruit γ-TuRCs to organize MTs and support spermiogenesis. This molecular characterization of the mitochondrial MTOC defines a minimal molecular requirement for MTOC generation and implicates the potent role of Cnn (or its related) proteins in the direct regulation of MT assembly and organization of non-centrosomal MTOCs.

Full length RTN3 regulates turnover of tubular endoplasmic reticulum via selective autophagy.

  • Grumati P
  • Elife
  • 2017 Jun 15

Literature context: (RRID:CVCL_0023) were prov


Abstract:

The turnover of endoplasmic reticulum (ER) ensures the correct biological activity of its distinct domains. In mammalian cells, the ER is degraded via a selective autophagy pathway (ER-phagy), mediated by two specific receptors: FAM134B, responsible for the turnover of ER sheets and SEC62 that regulates ER recovery following stress. Here, we identified reticulon 3 (RTN3) as a specific receptor for the degradation of ER tubules. Oligomerization of the long isoform of RTN3 is sufficient to trigger fragmentation of ER tubules. The long N-terminal region of RTN3 contains several newly identified LC3-interacting regions (LIR). Binding to LC3s/GABARAPs is essential for the fragmentation of ER tubules and their delivery to lysosomes. RTN3-mediated ER-phagy requires conventional autophagy components, but is independent of FAM134B. None of the other reticulon family members have the ability to induce fragmentation of ER tubules during starvation. Therefore, we assign a unique function to RTN3 during autophagy.

Systematic Quantification of Population Cell Death Kinetics in Mammalian Cells.

  • Forcina GC
  • Cell Syst
  • 2017 Jun 28

Literature context: CCL-185; RRID:CVCL_0023 Bax+/+Bak+


Abstract:

Cytotoxic compounds are important drugs and research tools. Here, we introduce a method, scalable time-lapse analysis of cell death kinetics (STACK), to quantify the kinetics of compound-induced cell death in mammalian cells at the population level. STACK uses live and dead cell markers, high-throughput time-lapse imaging, and mathematical modeling to determine the kinetics of population cell death over time. We used STACK to profile the effects of over 1,800 bioactive compounds on cell death in two human cancer cell lines, resulting in a large and freely available dataset. 79 potent lethal compounds common to both cell lines caused cell death with widely divergent kinetics. 13 compounds triggered cell death within hours, including the metallophore zinc pyrithione. Mechanistic studies demonstrated that this rapid onset lethal phenotype was caused in human cancer cells by metabolic disruption and ATP depletion. These results provide the first comprehensive survey of cell death kinetics and analysis of rapid-onset lethal compounds.

Funding information:
  • NCI NIH HHS - R00 CA166517()

The RNA Exosome Syncs IAV-RNAPII Transcription to Promote Viral Ribogenesis and Infectivity.

  • Rialdi A
  • Cell
  • 2017 May 4

Literature context: 7Experimental Models: Cell LinesA549ATCCCCL-185293TATCCCRL-3216MDCKA


Abstract:

The nuclear RNA exosome is an essential multi-subunit complex that controls RNA homeostasis. Congenital mutations in RNA exosome genes are associated with neurodegenerative diseases. Little is known about the role of the RNA exosome in the cellular response to pathogens. Here, using NGS and human and mouse genetics, we show that influenza A virus (IAV) ribogenesis and growth are suppressed by impaired RNA exosome activity. Mechanistically, the nuclear RNA exosome coordinates the initial steps of viral transcription with RNAPII at host promoters. The viral polymerase complex co-opts the nuclear RNA exosome complex and cellular RNAs en route to 3' end degradation. Exosome deficiency uncouples chromatin targeting of the viral polymerase complex and the formation of cellular:viral RNA hybrids, which are essential RNA intermediates that license transcription of antisense genomic viral RNAs. Our results suggest that evolutionary arms races have shaped the cellular RNA quality control machinery.

Funding information:
  • NIAID NIH HHS - R01 AI099195()
  • NIAID NIH HHS - U19 AI106754()
  • NIH HHS - DP2 OD008651()

RPL10L Is Required for Male Meiotic Division by Compensating for RPL10 during Meiotic Sex Chromosome Inactivation in Mice.

  • Jiang L
  • Curr. Biol.
  • 2017 May 22

Literature context: CCL-185; RRID:CVCL_0023 HeLa ATCC


Abstract:

The mammalian sex chromosomes have undergone profound changes during their evolution from an ancestral pair of autosomes [1-4]. Specifically, the X chromosome has acquired a paradoxical sex-biased function by redistributing gene contents [5, 6] and has generated a disproportionately high number of retrogenes that are located on autosomes and exhibit male-biased expression patterns [6]. Several selection-based models have been proposed to explain this phenomenon, including a model of sexual antagonism driving X inactivation (SAXI) [6-8] and a compensatory mechanism based on meiotic sex chromosome inactivation (MSCI) [6, 8-11]. However, experimental evidence correlating the function of X-chromosome-derived autosomal retrogenes with evolutionary forces remains limited [12-17]. Here, we show that the deficiency of Rpl10l, a murine autosomal retrogene of Rpl10 with testis-specific expression, disturbs ribosome biogenesis in late-prophase spermatocytes and prohibits the transition from prophase into metaphase of the first meiotic division, resulting in male infertility. Rpl10l expression compensates for the lack of Rpl10, which exhibits a broad expression pattern but is subject to MSCI during spermatogenesis. Importantly, ectopic expression of RPL10L prevents the death of cultured RPL10-deficient somatic cells, and Rpl10l-promoter-driven transgenic expression of Rpl10 in spermatocytes restores spermatogenesis and fertility in Rpl10l-deficient mice. Our results demonstrate that Rpl10l plays an essential role during the meiotic stage of spermatogenesis by compensating for MSCI-mediated transcriptional silencing of Rpl10. These data provide direct evidence for the compensatory hypothesis and add novel insight into the evolution of X-chromosome-derived autosomal retrogenes and their role in male fertility.

A Compendium of RNA-Binding Proteins that Regulate MicroRNA Biogenesis.

  • Treiber T
  • Mol. Cell
  • 2017 Apr 20

Literature context: ATCC® CCL-2human: A549ATCCATCC® CCL-185human: SK-MEL-28ATCCATCC® HTB-72


Abstract:

During microRNA (miRNA) biogenesis, two endonucleolytic reactions convert stem-loop-structured precursors into mature miRNAs. These processing steps can be posttranscriptionally regulated by RNA-binding proteins (RBPs). Here, we have used a proteomics-based pull-down approach to map and characterize the interactome of a multitude of pre-miRNAs. We identify ∼180 RBPs that interact specifically with distinct pre-miRNAs. For functional validation, we combined RNAi and CRISPR/Cas-mediated knockout experiments to analyze RBP-dependent changes in miRNA levels. Indeed, a large number of the investigated candidates, including splicing factors and other mRNA processing proteins, have effects on miRNA processing. As an example, we show that TRIM71/LIN41 is a potent regulator of miR-29a processing and its inactivation directly affects miR-29a targets. We provide an extended database of RBPs that interact with pre-miRNAs in extracts of different cell types, highlighting a widespread layer of co- and posttranscriptional regulation of miRNA biogenesis.

O2⋅- and H2O2-Mediated Disruption of Fe Metabolism Causes the Differential Susceptibility of NSCLC and GBM Cancer Cells to Pharmacological Ascorbate.

  • Schoenfeld JD
  • Cancer Cell
  • 2017 Apr 10

Literature context: ollection RRID:CVCL_0023 H1299 Amer


Abstract:

Pharmacological ascorbate has been proposed as a potential anti-cancer agent when combined with radiation and chemotherapy. The anti-cancer effects of ascorbate are hypothesized to involve the autoxidation of ascorbate leading to increased steady-state levels of H2O2; however, the mechanism(s) for cancer cell-selective toxicity remain unknown. The current study shows that alterations in cancer cell mitochondrial oxidative metabolism resulting in increased levels of O2⋅- and H2O2 are capable of disrupting intracellular iron metabolism, thereby selectively sensitizing non-small-cell lung cancer (NSCLC) and glioblastoma (GBM) cells to ascorbate through pro-oxidant chemistry involving redox-active labile iron and H2O2. In addition, preclinical studies and clinical trials demonstrate the feasibility, selective toxicity, tolerability, and potential efficacy of pharmacological ascorbate in GBM and NSCLC therapy.

Funding information:
  • NCI NIH HHS - P30 CA086862()
  • NCI NIH HHS - R01 CA169046()
  • NCI NIH HHS - R01 CA182804()
  • NCI NIH HHS - R01 CA184051()
  • NCI NIH HHS - T32 CA078586()
  • NCI NIH HHS - U01 CA140206()
  • NCI NIH HHS - U01 CA166800()
  • NIGMS NIH HHS - T32 GM007337()

p27Kip1 promotes invadopodia turnover and invasion through the regulation of the PAK1/Cortactin pathway.

  • Jeannot P
  • Elife
  • 2017 Mar 13

Literature context: and A549 (RRID:CVCL_0023) cells wer


Abstract:

p27Kip1 (p27) is a cyclin-CDK inhibitor and negative regulator of cell proliferation. p27 also controls other cellular processes including migration and cytoplasmic p27 can act as an oncogene. Furthermore, cytoplasmic p27 promotes invasion and metastasis, in part by promoting epithelial to mesenchymal transition. Herein, we find that p27 promotes cell invasion by binding to and regulating the activity of Cortactin, a critical regulator of invadopodia formation. p27 localizes to invadopodia and limits their number and activity. p27 promotes the interaction of Cortactin with PAK1. In turn, PAK1 promotes invadopodia turnover by phosphorylating Cortactin, and expression of Cortactin mutants for PAK-targeted sites abolishes p27's effect on invadopodia dynamics. Thus, in absence of p27, cells exhibit increased invadopodia stability due to impaired PAK1-Cortactin interaction, but their invasive capacity is reduced compared to wild-type cells. Overall, we find that p27 directly promotes cell invasion by facilitating invadopodia turnover via the Rac1/PAK1/Cortactin pathway.

Suppressor of cytokine signaling (SOCS)5 ameliorates influenza infection via inhibition of EGFR signaling.

  • Kedzierski L
  • Elife
  • 2017 Feb 14

Literature context: thod (Pierce). A549 cells (RRID:CVCL_0023) were treated with 10 μM MG132


Abstract:

Influenza virus infections have a significant impact on global human health. Individuals with suppressed immunity, or suffering from chronic inflammatory conditions such as COPD, are particularly susceptible to influenza. Here we show that suppressor of cytokine signaling (SOCS) five has a pivotal role in restricting influenza A virus in the airway epithelium, through the regulation of epidermal growth factor receptor (EGFR). Socs5-deficient mice exhibit heightened disease severity, with increased viral titres and weight loss. Socs5 levels were differentially regulated in response to distinct influenza viruses (H1N1, H3N2, H5N1 and H11N9) and were reduced in primary epithelial cells from COPD patients, again correlating with increased susceptibility to influenza. Importantly, restoration of SOCS5 levels restricted influenza virus infection, suggesting that manipulating SOCS5 expression and/or SOCS5 targets might be a novel therapeutic approach to influenza.

MK-8776, a novel chk1 kinase inhibitor, radiosensitizes p53-defective human tumor cells.

  • Bridges KA
  • Oncotarget
  • 2016 Nov 1

Literature context: CCL-185, RRID:CVCL_0023), H1299 (A


Abstract:

Radiotherapy is commonly used to treat a variety of solid tumors but improvements in the therapeutic ratio are sorely needed. The aim of this study was to assess the Chk1 kinase inhibitor, MK-8776, for its ability to radiosensitize human tumor cells. Cells derived from NSCLC and HNSCC cancers were tested for radiosensitization by MK-8776. The ability of MK-8776 to abrogate the radiation-induced G2 block was determined using flow cytometry. Effects on repair of radiation-induced DNA double strand breaks (DSBs) were determined on the basis of rad51, γ-H2AX and 53BP1 foci. Clonogenic survival analyses indicated that MK-8776 radiosensitized p53-defective tumor cells but not lines with wild-type p53. Abrogation of the G2 block was evident in both p53-defective cells and p53 wild-type lines indicating no correlation with radiosensitization. However, only p53-defective cells entered mitosis harboring unrepaired DSBs. MK-8776 appeared to inhibit repair of radiation-induced DSBs at early times after irradiation. A comparison of MK-8776 to the wee1 inhibitor, MK-1775, suggested both similarities and differences in their activities. In conclusion, MK-8776 radiosensitizes tumor cells by mechanisms that include abrogation of the G2 block and inhibition of DSB repair. Our findings support the clinical evaluation of MK-8776 in combination with radiation.

Funding information:
  • NCI NIH HHS - P30 CA016672()
  • NCI NIH HHS - R01 CA168485()

Human Adaptation of Ebola Virus during the West African Outbreak.

  • Urbanowicz RA
  • Cell
  • 2016 Nov 3

Literature context: 977 A549; RRID:CVCL_0023 Human hepa


Abstract:

The 2013-2016 outbreak of Ebola virus (EBOV) in West Africa was the largest recorded. It began following the cross-species transmission of EBOV from an animal reservoir, most likely bats, into humans, with phylogenetic analysis revealing the co-circulation of several viral lineages. We hypothesized that this prolonged human circulation led to genomic changes that increased viral transmissibility in humans. We generated a synthetic glycoprotein (GP) construct based on the earliest reported isolate and introduced amino acid substitutions that defined viral lineages. Mutant GPs were used to generate a panel of pseudoviruses, which were used to infect different human and bat cell lines. These data revealed that specific amino acid substitutions in the EBOV GP have increased tropism for human cells, while reducing tropism for bat cells. Such increased infectivity may have enhanced the ability of EBOV to transmit among humans and contributed to the wide geographic distribution of some viral lineages.

Funding information:
  • Medical Research Council - G0801169()

Regulation of mTORC1 by lysosomal calcium and calmodulin.

  • Li RJ
  • Elife
  • 2016 Oct 27

Literature context: and A549 (RRID:CVCL_0023, purchased


Abstract:

Blockade of lysosomal calcium release due to lysosomal lipid accumulation has been shown to inhibit mTORC1 signaling. However, the mechanism by which lysosomal calcium regulates mTORC1 has remained undefined. Herein we report that proper lysosomal calcium release through the calcium channel TRPML1 is required for mTORC1 activation. TRPML1 depletion inhibits mTORC1 activity, while overexpression or pharmacologic activation of TRPML1 has the opposite effect. Lysosomal calcium activates mTORC1 by inducing association of calmodulin (CaM) with mTOR. Blocking the interaction between mTOR and CaM by antagonists of CaM significantly inhibits mTORC1 activity. Moreover, CaM is capable of stimulating the kinase activity of mTORC1 in a calcium-dependent manner in vitro. These results reveal that mTOR is a new type of CaM-dependent kinase, and TRPML1, lysosomal calcium and CaM play essential regulatory roles in the mTORC1 signaling pathway.

Funding information:
  • NCATS NIH HHS - UL1 TR001079()
  • NCI NIH HHS - R01 CA184103()

TP53 exon-6 truncating mutations produce separation of function isoforms with pro-tumorigenic functions.

  • Shirole NH
  • Elife
  • 2016 Oct 19

Literature context: Mycoplasma contamination. A549 (RRID: CVCL_0023), AU565 (RRID: CVCL_1074), H129


Abstract:

TP53 truncating mutations are common in human tumors and are thought to give rise to p53-null alleles. Here, we show that TP53 exon-6 truncating mutations occur at higher than expected frequencies and produce proteins that lack canonical p53 tumor suppressor activities but promote cancer cell proliferation, survival, and metastasis. Functionally and molecularly, these p53 mutants resemble the naturally occurring alternative p53 splice variant, p53-psi. Accordingly, these mutants can localize to the mitochondria where they promote tumor phenotypes by binding and activating the mitochondria inner pore permeability regulator, Cyclophilin D (CypD). Together, our studies reveal that TP53 exon-6 truncating mutations, contrary to current beliefs, act beyond p53 loss to promote tumorigenesis, and could inform the development of strategies to target cancers driven by these prevalent mutations.

Funding information:
  • NCI NIH HHS - F31 CA192835()
  • NCI NIH HHS - P01 CA129243()
  • NCI NIH HHS - P30 CA008748()

Inactivation of oncogenic cAMP-specific phosphodiesterase 4D by miR-139-5p in response to p53 activation.

  • Cao B
  • Elife
  • 2016 Jul 7

Literature context: es. A549 (RRID:CVCL_0023), HepG2 (R


Abstract:

Increasing evidence highlights the important roles of microRNAs in mediating p53's tumor suppression functions. Here, we report miR-139-5p as another new p53 microRNA target. p53 induced the transcription of miR-139-5p, which in turn suppressed the protein levels of phosphodiesterase 4D (PDE4D), an oncogenic protein involved in multiple tumor promoting processes. Knockdown of p53 reversed these effects. Also, overexpression of miR-139-5p decreased PDE4D levels and increased cellular cAMP levels, leading to BIM-mediated cell growth arrest. Furthermore, our analysis of human colorectal tumor specimens revealed significant inverse correlation between the expression of miR-139-5p and that of PDE4D. Finally, overexpression of miR-139-5p suppressed the growth of xenograft tumors, accompanied by decrease in PDE4D and increase in BIM. These results demonstrate that p53 inactivates oncogenic PDE4D by inducing the expression of miR-139-5p.

Funding information:
  • NCI NIH HHS - R01 CA095441()
  • NCI NIH HHS - R01 CA127724()
  • NCI NIH HHS - R01 CA172468()
  • NCI NIH HHS - R21 CA190775()

Interferon-Induced Transmembrane Protein 3 Inhibits Hantaan Virus Infection, and Its Single Nucleotide Polymorphism rs12252 Influences the Severity of Hemorrhagic Fever with Renal Syndrome.

  • Xu-Yang Z
  • Front Immunol
  • 2016 Jan 18

Literature context: -CCL-185, RRID:CVCL_0023) were grow


Abstract:

Hantaan virus (HTNV) causes hemorrhagic fever with renal syndrome (HFRS). Previous studies have identified interferon-induced transmembrane proteins (IFITMs) as an interferon-stimulated gene family. However, the role of IFITMs in HTNV infection is unclear. In this study, we observed that IFITM3 single nucleotide polymorphisms (SNP) rs12252 C allele and CC genotype associated with the disease severity and HTNV load in the plasma of HFRS patients. In vitro experiments showed that the truncated protein produced by the rs12252 C allele exhibited an impaired anti-HTNV activity. We also proved that IFITM3 was able to inhibit HTNV infection in both HUVEC and A549 cells by overexpression and RNAi assays, likely via a mechanism of inhibiting virus entry demonstrated by binding and entry assay. Localization of IFITM3 in late endosomes was also observed. In addition, we demonstrated that the transcription of IFITM3 is negatively regulated by an lncRNA negative regulator of interferon response (NRIR). Taken together, we conclude that IFITM3, negatively regulated by NRIR, inhibits HTNV infection, and its SNP rs12252 correlates with the plasma HTNV load and the disease severity of patients with HFRS.