Abstract：
:We report the first RNAi-induced phenotypes in mammalian cultured cells using RNA interference mediated by duplexes of 21-nt RNAs. The 21 gene products studied have different functions and subcellular localizations. Knockdown experiments monitored by immunofluorescence and immunoblotting show that even major cellular proteins such as actin and vimentin can be silenced efficiently. Genes were classified as essential or nonessential depending on impaired cell growth after RNA silencing. Phenotypes also involved altered cell morphology and aberrant mitotic arrest. Among the essential genes identified by RNAi for which such information was previously not available are lamin B1, lamin B2, NUP153, GAS41, ARC21, cytoplasmic dynein, the protein kinase cdk1 and both beta- and gamma-actin. Newly defined nonessential genes are emerin and zyxin. Several genes previously characterized by other methods such as knockout of murine genes are included as internal controls and gave identical results when RNAi was used. In the case of two nonessential genes (lamin A/C and zyxin) RNAi provides a recognizable phenotype. Our results complete the characterization of the mammalian nuclear lamins. While lamins A/C appear as nonessential proteins in the mouse embryo and in RNAi treated cultured cells, the two other lamins, B1 and B2, are now identified as essential proteins. Interestingly the inner nuclear membrane protein emerin, thought to be a ligand of lamin A/C, is also a nonessential protein in tissue culture cells.
journal_name
J Cell Scijournal_title
Journal of cell scienceauthors
Harborth J,Elbashir SM,Bechert K,Tuschl T,Weber Kkeywords:
subject
Has Abstractpub_date
2001-12-01 00:00:00pages
4557-65issue
Pt 24eissn
0021-9533issn
1477-9137journal_volume
114pub_type
杂志文章abstract：:Autophagy requires the formation of membrane vesicles, known as autophagosomes, that engulf cellular cargoes and subsequently recruit lysosomal hydrolases for the degradation of their contents. A number of autophagy-related proteins act to mediate the de novo biogenesis of autophagosomes and vesicular trafficking even...
journal_title：Journal of cell science
pub_type： 杂志文章,评审
doi：10.1242/jcs.249227
更新日期：2020-10-30 00:00:00
abstract：:Microdomains have been proposed to explain specificity in the myriad of possible cellular targets of cAMP. Local differences in cAMP levels can be generated by phosphodiesterases, which control the diffusion of cAMP. Here, we address the possibility that adenylyl cyclases, the source of cAMP, can be primary architects...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.058594
更新日期：2010-01-01 00:00:00
abstract：:The attachment of polymorphonuclear leukocytes (PMN) to the endothelial lining of blood vessels is an important initial event in the acute inflammatory reaction. Experiments were carried out to examine some of the parameters of this adhesive interaction and the extent to which the substrate (endothelium) plays a role ...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1980-10-01 00:00:00
abstract：:The Na(+)/H(+) exchanger NHE1 is involved in intracellular pH homeostasis and cell volume regulation and accumulates with actin in the lamellipodia of fibroblasts. In order to determine the role of NHE1 following epithelial transformation and the acquisition of motile and invasive properties, we studied NHE1 expressio...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：2000-10-01 00:00:00
abstract：:Aquaporin-4, the major water-selective channel in astroglia throughout the central nervous system, facilitates water movement into and out of the brain. Here, we identify a novel role for aquaporin-4 in astroglial cell migration, as occurs during glial scar formation. Astroglia cultured from the neocortex of aquaporin...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.02680
更新日期：2005-12-15 00:00:00
abstract：:Our previous studies of PAWS1 (protein associated with SMAD1; also known as FAM83G) have suggested that this molecule has roles beyond BMP signalling. To investigate these roles, we have used CRISPR/Cas9 to generate PAWS1-knockout U2OS osteosarcoma cells. Here, we show that PAWS1 plays a role in the regulation of the ...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.202390
更新日期：2018-01-10 00:00:00
abstract：:Ca(2+) signaling mediated by phospholipase C that produces inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] and diacylglycerol (DAG) controls lymphocyte activation. In contrast to store-operated Ca(2+) entry activated by Ins(1,4,5)P(3)-induced Ca(2+) release from endoplasmic reticulum, the importance of DAG-activated Ca(...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.061051
更新日期：2010-03-15 00:00:00
abstract：:Euglena gracilis is a suitable model system to investigate the role of zinc in the process of cell division. In zinc-deficient organisms there is a characteristic arrest of cellular proliferation, the DNA content of the cells doubles, whereas RNA and protein contents decrease. The present investigations include the gr...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1975-01-01 00:00:00
abstract：:Adhesion between cells is established by the formation of specialized intercellular junctional complexes, such as desmosomes. Desmosomes contain isoforms of two members of the cadherin superfamily of cell adhesion proteins, desmocollins (Dsc) and desmogleins (Dsg), but their combinatorial roles in desmosome assembly a...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.146316
更新日期：2014-05-15 00:00:00
abstract：:Postsynaptic density protein 95 (PSD-95/SAP-90) is a palmitoylated membrane-associated guanylate kinase that oligomerizes and clusters ion channels and associated signaling machinery at excitatory synapses in brain. However, the mechanism for PSD-95 oligomerization and its relationship to ion channel clustering remain...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.00617
更新日期：2003-08-01 00:00:00
abstract：:The non-alpha-helical N-terminal domain of intermediate filament proteins plays a key role in filament assembly. Previous studies have identified a nonapeptide motif, SSYRRIFGG, in the non-alpha-helical N-terminal domain of vimentin that is required for assembly. This motif is also found in desmin, peripherin and the ...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1994-07-01 00:00:00
abstract：:Previous studies have shown that the non-alpha-helical, amino-terminal head region of vimentin is essential for the formation and stability of vimentin intermediate filaments (IFs). In order to specify its target site on companion protein subunits, it was cut off from vimentin at amino acid position 96 with lysine-spe...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1992-02-01 00:00:00
abstract：:Apical and basolateral endocytic pathways in polarised Caco-2 cells were investigated by following the uptake, recycling and transcytosis of the galactose-binding protein toxin ricin, as a membrane marker. Differences in the extent and kinetics of lectin uptake, recycling and transcytosis were observed at the apical a...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1994-09-01 00:00:00
abstract：:We have examined the pattern of expression of four different matrix metalloproteinases (MMPs), collagenase, stromelysin, 92 kD gelatinase, and 72 kD gelatinase, by primary and passaged cultures of rabbit corneal fibroblasts. Primary cultures of this cell type have previously been shown to reproduce the normal tissue r...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1990-10-01 00:00:00
abstract：:Epithelial cell-cell junctions are specialised structures connecting individual cells in epithelial tissues. They are dynamically and functionally linked to the actin cytoskeleton. Disassembly of these junctions is a key event during physiological and pathological processes, but how this influences gene expression is ...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.014456
更新日期：2008-04-01 00:00:00
abstract：:Recent studies suggest that cytochrome P450 (CYP) epoxygenase-derived epoxyeicosatrienoic acids (EETs) elicit cell proliferation and promote angiogenesis. The aim of this study was to determine the role of CYP 2C8/9-derived EETs in the process of angiogenesis under hypoxic conditions. In human endothelial cells, hypox...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.02674
更新日期：2005-12-01 00:00:00
abstract：:T cell activation is accompanied by actin-mediated receptor clustering and reorganization of lipid rafts. It has been suggested that costimulatory molecules might be involved in these processes. We here provide evidence that engagement of the adhesion molecule CD44 initiates cytoskeletal rearrangement and membrane reo...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：2001-03-01 00:00:00
abstract：:The tumor suppressor p53 is activated in response to many forms of cellular stress leading to cell cycle arrest, senescence or apoptosis. Appropriate sub-cellular localization is essential for modulating p53 function. We recently showed that p53 localizes to the nucleolus after proteasome inhibition with MG132 and thi...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.030098
更新日期：2008-12-15 00:00:00
abstract：:Quantitative information about the range of influence of extracellular signaling molecules is critical for understanding their effects, but is difficult to determine in the complex and dynamic three-dimensional environment of a living embryo. Drosophila germ cells migrate during embryogenesis and use spatial informati...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.136176
更新日期：2013-12-01 00:00:00
abstract：:PML nuclear bodies (NBs) are involved in the regulation of key nuclear pathways but their biochemical function in nuclear metabolism is unknown. In this study PML NB assembly dynamics were assessed by live cell imaging and mathematic modeling of its major component parts. We show that all six nuclear PML isoforms exhi...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.031922
更新日期：2008-08-15 00:00:00
abstract：:We report a fast method for the isolation of homogeneous C-protein from bovine skeletal muscle. In electron micrographs C-protein appears as short rods with a relatively uniform length of about 50 nm. Protein sequencing shows a single N-terminal sequence. Radio-labelled C-protein strongly decorates titin II and myosin...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1992-08-01 00:00:00
abstract：:In metazoans, cyclin-dependent kinase 7 (CDK7) has essential roles in both the cell-division cycle and transcription, as a CDK-activating kinase (CAK) and as a component of the general transcription factor TFIIH, respectively. Controversy over its double duty has been resolved, but questions remain. First, how does CD...
journal_title：Journal of cell science
pub_type： 杂志文章,评审
doi：10.1242/jcs.02718
更新日期：2005-11-15 00:00:00
abstract：:Patients with myotonic dystrophy type 1 exhibit a diversity of symptoms that affect many different organs. Among these are cognitive dysfunctions, the origin of which has remained elusive, partly because of the difficulty in accessing neural cells. Here, we have taken advantage of pluripotent stem cell lines derived f...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.116285
更新日期：2013-04-15 00:00:00
abstract：:Intracellular signal transduction occurs through cascades of reactions involving dozens of proteins that transmit signals from the cell surface, through a crowded cellular environment filled with organelles and a filamentous cytoskeleton, to specific targets. Numerous signaling molecules are immobilized or transiently...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.01122
更新日期：2004-06-01 00:00:00
abstract：:Cytostellin, a approximately 240 kDa phosphoprotein found in all cells examined from human to yeast, is predominantly intranuclear in interphase mammalian cells and undergoes continuous redistribution during the cell cycle. Here, mammalian cytostellin is shown to localize to intranuclear regions enriched with multiple...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1994-03-01 00:00:00
abstract：:Vertebrate tropomyosins (TMs) are expressed from four genes, and at least 18 distinct isoforms are generated via a complex pattern of alternative RNA splicing and alternative promoters. The functional significance of this isoform diversity is largely unknown and it remains to be determined whether specific isoforms ar...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1999-02-01 00:00:00
abstract：:Endocytosis of chemokine receptors regulates signal transduction initiated by chemokines, but the molecular mechanisms underlying this process are not fully defined. In this work, we assessed the involvement of the motor protein nonmuscle myosin heavy chain IIA (MIIA) in the endocytosis of CXCR4 induced by SDF-1alpha ...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：10.1242/jcs.03415
更新日期：2007-03-15 00:00:00
abstract：:Tau is a neuronal microtubule-associated protein that appears to function in the formation and maintenance of axons by influencing microtubule organisation. Tau is a phosphoprotein and is more heavily phosphorylated in fetal than in adult brain, and is also hyperphosphorylated in Alzheimer's disease where it forms the...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1996-06-01 00:00:00
abstract：:Tenascin, an extracellular matrix protein that modulates cell adhesion, exists as a unique six-armed structure called a hexabrachion. The human hexabrachion is composed of six identical 320 kDa subunits and the structure is stabilized by inter-subunit disulfide bonds between amino-terminal segments. We have examined t...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1995-04-01 00:00:00
abstract：:Phagosomes are formed when phagocytic cells ingest particles such as bacteria, viruses or synthetic beads of different kinds. The environment within the phagosome gradually changes to generate degradative conditions. These changes require multiple interactions between the maturing phagosomes and the endocytic and the ...
journal_title：Journal of cell science
pub_type： 杂志文章
doi：
更新日期：1998-01-01 00:00:00
