Abstract:
:The discovery of fluorescent proteins (FPs) has revolutionized cell biology. The fusion of targeting sequences to FPs enables the investigation of cellular organelles and their dynamics; however, occasionally, such fluorescent fusion proteins (FFPs) exhibit behavior different from that of the native proteins. Here, we constructed a color pallet comprising different organelle markers and found that FFPs targeted to the mitochondria were mislocalized when fused to certain types of FPs. Such FPs included several variants of Aequorea victoria green FP (avGFP) and a monomeric variant of the red FP. Because the FFPs that are mislocalized include FPs with faster maturing or folding mutations, the increase in the maturation rate is likely to prevent their expected localization. Indeed, when we reintroduced amino acid substitutions so that the FP sequences were equivalent to that of wild-type avGFP, FFP localization to the mitochondria was significantly enhanced. Moreover, similar amino acid substitutions improved the localization of mitochondria-targeted pHluorin, which is a pH-sensitive variant of GFP, and its capability to monitor pH changes in the mitochondrial matrix. Our findings demonstrate the importance of selecting FPs that maximize FFP function.Key words: fluorescent protein, organelle, fusion protein, mitochondria.
journal_name
Cell Struct Functjournal_title
Cell structure and functionauthors
Kashiwagi S,Fujioka Y,Satoh AO,Yoshida A,Fujioka M,Nepal P,Tsuzuki A,Aoki O,Paudel S,Sasajima H,Ohba Ydoi
10.1247/csf.19028subject
Has Abstractpub_date
2019-12-26 00:00:00pages
183-194issue
2eissn
0386-7196issn
1347-3700journal_volume
44pub_type
杂志文章abstract::Significant progress has been made toward our understanding of the basic principle of nucleocytoplasmic transport, and the structure of transport factors, as well as the diversity of nucleocytoplasmic transport pathways. This review outlines the current knowledge of transport, and discusses the problems that remain as...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.25.207
更新日期:2000-08-01 00:00:00
abstract::To elucidate the cytotoxic mechanism of tumor necrosis factor (TNF), we isolated TNF-resistant sublines of L929 cells. As compared with L929 cells, TNF-resistant cells retained similar number and affinity of TNF-binding sites, and showed a similar growth rate. TNF stimulated arachidonate release from L929 cells, while...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.16.333
更新日期:1991-08-01 00:00:00
abstract::Histone modifications play critical roles in the epigenetic regulation of gene expression and in the maintenance of genome integrity. Acetylation and methylation of histone H3 are particularly important in gene activation and silencing. We generated and characterized a panel of mouse monoclonal antibodies that specifi...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.07035
更新日期:2008-01-01 00:00:00
abstract::In view of the importance of vitamin A in the human immune system and the central role of interleukin-2 (IL-2) in the proliferation of T-lymphocytes, we examined the effects of all-trans-retinoic acid (ATRA) on the protein and gene expression of IL-2 in the human T-cell line HUT-78 when stimulated with either 12-O-tet...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.07020
更新日期:2008-01-01 00:00:00
abstract::We earlier discovered a novel 40-kDa protein (hsp40) induced by heat shock and other stresses in mammalian and avian cells. In this report, we purified the hsp40 in HeLa cells, using modified two-dimensional gel electrophoresis, and determined the amino terminal amino acid sequence of this protein. The hsp40 is homolo...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.17.77
更新日期:1992-02-01 00:00:00
abstract::Previously, we serially transplanted tumors that produced colony-stimulating factor (CSF) into nude mice, who developed marked granulocytosis along with tumor growth; their leukocyte counts reaching approximately one million per cu mm. The numbers of CFU-GM, CFU-E, CFU-Meg, CFU-S and BFU-E were increased in nude mice ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.12.273
更新日期:1987-06-01 00:00:00
abstract::Recent studies have demonstrated that vesicle transport to cleavage furrow is indispensable for cytokinesis. Some animal and plant cells form distinct structures during cell division known as central spindle and phragmoplast, respectively. Several essential factors involved in the vesicle transport have been isolated ...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.26.567
更新日期:2001-12-01 00:00:00
abstract::Newly prepared phosphatidylethanolamine (PE) conjugates of glycosaminoglycans (GAGs) enabled us to immobilize GAGs to solid phase through hydrophobic interaction. Using these compounds, called GAG-PEs, we studied the effects of GAGs immobilized to culture plates coated with various extracellular matrix (ECM) proteins ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.20.199
更新日期:1995-06-01 00:00:00
abstract::alpha-1,3-Fucosyltransferase (Fuc T IV) cDNA was placed under the control of beta-actin, cytomegalovirus enhancer/promoter and transfected into embryonic stem cells. The transfected cell clones with integrated cDNA (positive clones) differentiated more efficiently into myocardial cell clones without integrated cDNA (n...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.22.247
更新日期:1997-04-01 00:00:00
abstract::Ran is a nuclear Ras-like GTPase that is required for various nuclear events including the bi-directional transport of proteins and ribonucleoproteins through the nuclear pore complex, spindle formation, and reassembly of the nuclear envelope. One of the key regulators of Ran is RanGAP1, a Ran specific GTPase activati...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.30.69
更新日期:2005-01-01 00:00:00
abstract::Hepatic stellate cells (HSCs), a mesenchymal cell type in hepatic parenchyma, have unique features with respect to their cellular origin, morphology, and function. Normal, quiescent HSCs function as major vitamin A-storing cells containing over 80% of total vitamin A in the body to maintain vitamin A homeostasis. HSCs...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.28.105
更新日期:2003-04-01 00:00:00
abstract::Contact reaction is the morphological expression of allogeneic recognition by hemocytes in the ascidian, Halocynthia roretzi; namely they undergo an exocytotic burst upon contact with non-self (allogeneic) hemocytes in vitro. We have found that contact reaction is accompanied by a burst of cyanide-insensitive oxygen c...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.20.81
更新日期:1995-02-01 00:00:00
abstract::NBT-II cells on collagen-coated substrates move rapidly and persistently, maintaining a semi-circular shape with a large lamellipodium, in a manner similar to fish keratocytes. The inhibitor of phospholipase D (PLD), n-butanol, completely blocked the migration and disturbed the characteristic localization of actin alo...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.07042
更新日期:2008-01-01 00:00:00
abstract::The putative ionophoretic action of phosphatidic acid or arachidonic acid metabolites for Ca2+ has offered an attractive explanation for stimulation-coupled mobilization of cytoplasmic Ca2+. We have examined the effects of Ca2+ ionophore and long-chain unsaturated fatty acids on the translocation of Ca2+ across the li...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.10.177
更新日期:1985-06-01 00:00:00
abstract::Genomics and proteomics studies in Chlamydomonas have revealed that an axoneme is composed of 200-600 types of proteins, including uncharacterized proteins collectively named flagellar-associated proteins (FAPs). Nine FAPs contain the EF-hand motif; however, they have not yet been well characterized. To find component...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.17023
更新日期:2018-02-16 00:00:00
abstract::Effects of kinase inhibitors on the preprophase band of microtubules in onion (Allium cepa L.) root tip cells were examined. Bundled microtubules in preprophase bands were dispersed on the cell cortex when onion seedlings were incubated with 2.5-5.0 mM 6-dimethylaminopurine. Fifteen min was enough for the bundled micr...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.24.419
更新日期:1999-10-01 00:00:00
abstract::The efficacy of anticancer agents significantly depends on the differential susceptibility of undifferentiated cancer cells and differentiated normal cells to undergo apoptosis. We previously found that enhanced expression of RPS3a/nbl, which apparently encodes a ribosomal protein, seems to prime cells for apoptosis, ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.25.103
更新日期:2000-04-01 00:00:00
abstract::Numerous types of cancer cells migrate into extracellular tissues. This phenomenon is termed invasion, and is associated with poor prognosis in cancer patients. In this study, we demonstrated that filamin B (FLNb), an actin-binding protein, is highly expressed in cancer cell lines that exhibit high invasiveness, with ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.15001
更新日期:2015-01-01 00:00:00
abstract::When spores of Dictyostelium discoideum matured through normal development were stained with rhodamine-phalloidin, rod-shaped fluorescence was found across the nucleus and in the cytoplasm. Electron micrographs of the rod shape showed that it consisted of long and straight fibers 12 nm in diameter. In contrast, a conv...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.19.189
更新日期:1994-08-01 00:00:00
abstract::The substance in sperm which induces the activation of eggs at fertilization has yet to be elucidated. Osawa et al. reported that the soluble extract of sperm (spex) causes inositol 1,4,5-trisphosphate (IP3)-induced Ca2+ release (IICR) in fertilized eggs of sea urchins when externally applied [Osawa et al., (1992). Zo...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.19.73
更新日期:1994-04-01 00:00:00
abstract::CD9 and CD63 belong to a tetramembrane-spanning glycoprotein family called tetraspanin, and are involved in a wide variety of cellular processes, but the structure-function relationship of this family of proteins has yet to be clarified. CD9 associates with diphtheria toxin receptor (DTR), which is identical to the me...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.25.317
更新日期:2000-10-01 00:00:00
abstract::Cultured fibroblasts from patients with Niemann-Pick disease type C (NP-C) are characterized by the lysosomal accumulation of unesterified cholesterol and the inability of low-density lipoprotein (LDL) to stimulate cholesterol esterification, in addition to impaired LDL-mediated down-regulation of LDL receptor activit...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.18.231
更新日期:1993-08-01 00:00:00
abstract::When human erythroleukemia cells (K562) were exposed to phorbol-12-myristate 13-acetate (PMA), phosphorylation of transferrin receptors was enhanced 5-fold with 10(-7) M PMA and 7-fold with 10(-6) M PMA, but not with 4 alpha-phorbol (5 X 10(-7) M). Stimulation took place in serine residues in the cytoplasmic domain of...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.10.95
更新日期:1985-06-01 00:00:00
abstract::The precise control of spindle microtubule (MT) dynamics is essential for chromosome capture and alignment. Kif18A/kinesin-8, an essential regulator of kinetochore MT dynamics, accumulates at its plus-ends in metaphase but not prometaphase cells. The underlying mechanism of time-dependent and kinetochore MT-specific p...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.11032
更新日期:2011-01-01 00:00:00
abstract::Microtubules are intrinsically dynamic polymers. Two kinds of dynamic behaviors, dynamic instability and treadmilling, are important for microtubule function in cells. Both dynamic behaviors appear to be tightly regulated, but the cellular molecules and the mechanisms responsible for the regulation remain largely unex...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.24.329
更新日期:1999-10-01 00:00:00
abstract::Under unfavorable conditions for growth, haploid myxoamoebae of Physarum polycephalum retracted their pseudopodia and changed their cell shape into disk-like form, after which they constructed the cell walls to form microcysts. These morphological changes of haploid cells were associated with changes in intracellular ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.17.301
更新日期:1992-10-01 00:00:00
abstract::The proto-oncogene c-myc is a multifunctional gene that regulates cell division, cell growth, and apoptosis. Here we report a new function of c-myc: induction of autophagy. Autophagy is a bulk degradation system for intracellular proteins. Autophagy proceeds with characteristic morphologies, which begins with the form...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.28.195
更新日期:2003-06-01 00:00:00
abstract::The Acanthamoeba myosin-IA and myosin-IB molecular motors bind to membranes, so they may produce the force to move organelles and membranes along actin filaments. We have determined the rate constants for the actin-activated myosin-I ATPase by pre-steady state kinetic analysis. ATP binds rapidly to myosin-I and dissoc...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.21.351
更新日期:1996-10-01 00:00:00
abstract::Here we report a new method of isolating epidermal desmosomes from Xenopus laevis, and a major constituent of desmosomes designated as Xenopus desmogleins (XDsg). Isolation of desmosomes from Xenopus laevis epidermis was carried out by a two step-incubation with different concentrations of NP-40. After discontinuous s...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.29.17
更新日期:2004-02-01 00:00:00
abstract::Pericentric regions form epigenetically organized, silent heterochromatin structures that accumulate histone H3 lysine 9 tri-methylation (H3K9me3) and heterochromatin protein 1 (HP1), a methylated H3K9-binding protein. At pericentric regions, Suv39h is the major enzyme that generates H3K9me3. Suv39h also interacts dir...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.16013
更新日期:2016-12-03 00:00:00