Golgi twins in late mitosis revealed by genetically encoded tags for live cell imaging and correlated electron microscopy.

Abstract:

:Combinations of molecular tags visible in light and electron microscopes become particularly advantageous in the analysis of dynamic cellular components like the Golgi apparatus. This organelle disassembles at the onset of mitosis and, after a sequence of poorly understood events, reassembles after cytokinesis. The precise location of Golgi membranes and resident proteins during mitosis remains unclear, partly due to limitations of molecular markers and the resolution of light microscopy. We generated a fusion consisting of the first 117 residues of alpha-mannosidase II tagged with a fluorescent protein and a tetracysteine motif. The mannosidase component guarantees docking into the Golgi membrane, with the tags exposed in the lumen. The fluorescent protein is optically visible without further treatment, whereas the tetracysteine tag can be reduced acutely with a membrane-permeant phosphine, labeled with ReAsH, monitored in the light microscope, and used to trigger the photoconversion of diaminobenzidine, allowing 4D optical recording on live cells and correlated ultrastructural analysis by electron microscopy. These methods reveal that Golgi reassembly is preceded by the formation of four colinear clusters at telophase, two per daughter cell. Within each daughter, the smaller cluster near the midbody gradually migrates to rejoin the major cluster on the far side of the nucleus and asymmetrically reconstitutes a single Golgi apparatus, first in one daughter cell and then in the other. Our studies provide previously undescribed insights into Golgi disassociation and reassembly during mitosis and offer a powerful approach to follow recombinant protein distribution in 4D imaging and correlated high-resolution analysis.

authors

Gaietta GM,Giepmans BN,Deerinck TJ,Smith WB,Ngan L,Llopis J,Adams SR,Tsien RY,Ellisman MH

doi

10.1073/pnas.0608509103

subject

Has Abstract

pub_date

2006-11-21 00:00:00

pages

17777-82

issue

47

eissn

0027-8424

issn

1091-6490

pii

0608509103

journal_volume

103

pub_type

杂志文章
  • Calmodulin priming: nuclear translocation of a calmodulin complex and the memory of prior neuronal activity.

    abstract::The neuronal nucleus plays a vital role in information processing, but whether it supports computational functions such as paired-pulse facilitation, comparable to synapses, is unclear. Ca(2+)-dependent movement of calmodulin (CaM) to the nucleus is highly responsive to Ca(2+) entry through L-type channels and promote...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.211563998

    authors: Mermelstein PG,Deisseroth K,Dasgupta N,Isaksen AL,Tsien RW

    更新日期:2001-12-18 00:00:00

  • Growth, metabolic partitioning, and the size of microorganisms.

    abstract::Population growth rate is a fundamental ecological and evolutionary characteristic of living organisms, but individuals must balance the metabolism devoted to biosynthesis and reproduction against the maintenance of existing structure and other functionality. Here we present a mathematical model that relates metabolic...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1115585109

    authors: Kempes CP,Dutkiewicz S,Follows MJ

    更新日期:2012-01-10 00:00:00

  • Regulation of intramitochondrial cholesterol transfer to side-chain cleavage cytochrome P-450 in rat adrenal gland.

    abstract::Rat adrenal mitochondria accumulated cholesterol during ether stress in vivo when side-chain cleavage was inhibited by aminoglutethimide (control = 14.6 vs. aminoglutethimide = 26.5 micrograms of cholesterol per mg of protein). This accumulation was insensitive to simultaneous administration of cycloheximide (24.2 mic...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.80.3.702

    authors: Privalle CT,Crivello JF,Jefcoate CR

    更新日期:1983-02-01 00:00:00

  • Monoclonal antibodies to the human insulin receptor block insulin binding and inhibit insulin action.

    abstract::Antibodies to the insulin receptor were prepared in BALB/c mice by immunization with IM-9 human lymphocytes, a cell type that has a large number of plasma membrane insulin receptors. The spleens of these mice were then removed, and their lymphocytes were fused to a mouse myeloma cell line, FO cells. After screening ov...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.79.23.7312

    authors: Roth RA,Cassell DJ,Wong KY,Maddux BA,Goldfine ID

    更新日期:1982-12-01 00:00:00

  • Membrane-bound MinDE complex acts as a toggle switch that drives Min oscillation coupled to cytoplasmic depletion of MinD.

    abstract::The Escherichia coli Min system self-organizes into a cell-pole to cell-pole oscillator on the membrane to prevent divisions at the cell poles. Reconstituting the Min system on a lipid bilayer has contributed to elucidating the oscillatory mechanism. However, previous in vitro patterns were attained with protein densi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1600644113

    authors: Vecchiarelli AG,Li M,Mizuuchi M,Hwang LC,Seol Y,Neuman KC,Mizuuchi K

    更新日期:2016-03-15 00:00:00

  • Reversal of malignant transformation by tumor DNA.

    abstract::Walker-256 carcinosarcoma cells grown in tissue culture medium containing inhibitory DNA prepared from the tumor are shown to have an unaltered growth in vitro, but a diminished capacity to produce malignant tumors on injection into the rat. Coincident with this change in virulence is the induction of tRNA methylase i...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.67.4.1827

    authors: Halpern BC,Halpern RM,Chaney SQ,Smith RA

    更新日期:1970-12-01 00:00:00

  • Colloquium paper: phylogenetic trees and the future of mammalian biodiversity.

    abstract::Phylogenies describe the origins and history of species. However, they can also help to predict species' fates and so can be useful tools for managing the future of biodiversity. This article starts by sketching how phylogenetic, geographic, and trait information can be combined to elucidate present mammalian diversit...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0801917105

    authors: Davies TJ,Fritz SA,Grenyer R,Orme CD,Bielby J,Bininda-Emonds OR,Cardillo M,Jones KE,Gittleman JL,Mace GM,Purvis A

    更新日期:2008-08-12 00:00:00

  • Sequence of the human 40-kDa keratin reveals an unusual structure with very high sequence identity to the corresponding bovine keratin.

    abstract::The complete amino acid and DNA sequences of the human 40-kDa keratin are reported. The DNA sequence encodes a protein of 44,098 Da, which is unique in that it lacks the terminal non-alpha-helical tail segment found in all other keratins. When the human 40-kDa keratin amino acid sequence is compared to the correspondi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.85.4.1114

    authors: Eckert RL

    更新日期:1988-02-01 00:00:00

  • Motion of a slightly compressible fluid.

    abstract::We show that the motion of a slightly compressible fluid is near that of an incompressible fluid. That is, for a given initial velocity field, the motion of a compressible fluid with large sound speed is near to that of an idealized incompressible fluid. We consider the compressible fluid motion in Lagrangian coordina...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.72.2.539

    authors: Ebin DG

    更新日期:1975-02-01 00:00:00

  • Transport of phosphocholine in higher plant cells: 31P nuclear magnetic resonance studies.

    abstract::Phosphocholine (PC) is an abundant primary form of organic phosphate that is transported in plant xylem sap. Addition of PC to the perfusate of compressed Pi-starved sycamore cells monitored by 31P NMR spectroscopy resulted in an accumulation of PC and all the other phosphate esters in the cytoplasmic compartment. Add...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.87.11.4280

    authors: Gout E,Bligny R,Roby C,Douce R

    更新日期:1990-06-01 00:00:00

  • Evidence for a genetic discontinuity between Neandertals and 24,000-year-old anatomically modern Europeans.

    abstract::During the late Pleistocene, early anatomically modern humans coexisted in Europe with the anatomically archaic Neandertals for some thousand years. Under the recent variants of the multiregional model of human evolution, modern and archaic forms were different but related populations within a single evolving species,...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1130343100

    authors: Caramelli D,Lalueza-Fox C,Vernesi C,Lari M,Casoli A,Mallegni F,Chiarelli B,Dupanloup I,Bertranpetit J,Barbujani G,Bertorelle G

    更新日期:2003-05-27 00:00:00

  • Loose coupling in the bacterial flagellar motor.

    abstract::Physiological properties of the flagellar rotary motor have been taken to indicate a tightly coupled mechanism in which each revolution is driven by a fixed number of energizing ions. Measurements that would directly test the tight-coupling hypothesis have not been made. Energizing ions flow through membrane-bound com...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1419955112

    authors: Boschert R,Adler FR,Blair DF

    更新日期:2015-04-14 00:00:00

  • Transforming neural computations and representing time.

    abstract::Motifs of neural circuitry seem surprisingly conserved over different areas of neocortex or of paleocortex, while performing quite different sensory processing tasks. This apparent paradox may be resolved by the fact that seemingly different problems in sensory information processing are related by transformations (ch...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.93.26.15440

    authors: Hopfield JJ

    更新日期:1996-12-24 00:00:00

  • Protein D1 preferentially binds A + T-rich DNA in vitro and is a component of Drosophila melanogaster nucleosomes containing A + T-rich satellite DNA.

    abstract::Our previous work [Levinger, L. & Varshavsky, A. (1982) Cell 28, 375-385] has shown that D1, a 50-kilodalton chromosomal protein of Drosophila melanogaster, is specifically associated with isolated nucleosomes that contain a complex A + T-rich satellite DNA with buoyant density of 1.688 g/ml. We show here that D1 is a...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.79.23.7152

    authors: Levinger L,Varshavsky A

    更新日期:1982-12-01 00:00:00

  • A regulator of nutritional copper signaling in Chlamydomonas is an SBP domain protein that recognizes the GTAC core of copper response element.

    abstract::The CRR1 (Copper Response Regulator) locus, required for both activating and repressing target genes of a copper- and hypoxia-sensing pathway in Chlamydomonas, encodes a 1,232-residue candidate transcription factor with a plant-specific DNA-binding domain named SBP, ankyrin repeats, and a C-terminal Cys-rich region, w...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0507693102

    authors: Kropat J,Tottey S,Birkenbihl RP,Depège N,Huijser P,Merchant S

    更新日期:2005-12-20 00:00:00

  • Pivotal role of amino acid at position 138 in the allosteric hinge reorientation of cAMP receptor protein.

    abstract::The cAMP receptor protein (CRP) of Escherichia coli needs cAMP for an allosteric change to regulate gene expression by binding to specific DNA sites. The hinge region connecting the DNA-binding domain to the cAMP-binding domain has been proposed to participate in the cAMP-induced allosteric change necessary to adjust ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.90.1.75

    authors: Ryu S,Kim J,Adhya S,Garges S

    更新日期:1993-01-01 00:00:00

  • Label-free imaging, detection, and mass measurement of single viruses by surface plasmon resonance.

    abstract::We report on label-free imaging, detection, and mass/size measurement of single viral particles in solution by high-resolution surface plasmon resonance microscopy. Diffraction of propagating plasmon waves along a metal surface by the viral particles creates images of the individual particles, which allow us to detect...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1005264107

    authors: Wang S,Shan X,Patel U,Huang X,Lu J,Li J,Tao N

    更新日期:2010-09-14 00:00:00

  • Cavbeta-subunit displacement is a key step to induce the reluctant state of P/Q calcium channels by direct G protein regulation.

    abstract::P/Q Ca(2+) channel activity is inhibited by G protein-coupled receptor activation. Channel inhibition requires a direct Gbetagamma binding onto the pore-forming subunit, Ca(v)2.1. It is characterized by biophysical changes, including current amplitude reduction, activation kinetic slowing, and an I-V curve shift, whic...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0306804101

    authors: Sandoz G,Lopez-Gonzalez I,Grunwald D,Bichet D,Altafaj X,Weiss N,Ronjat M,Dupuis A,De Waard M

    更新日期:2004-04-20 00:00:00

  • Selective loss of wheat germ agglutinin binding to agglutinin-resistant mutants of Chinese hamster ovary cells.

    abstract::The binding of 125I-labeled wheat germ agglutinin (WGA) to parental and three distinct WGA-resistant Chinese hamster ovary cell lines possessing modified cell surface carbohydrate structures has been examined over a 10(6)-fold range of WGA concentrations. The Scatchard plot for WGA binding to parental cells was comple...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.74.11.5056

    authors: Stanley P,Carver JP

    更新日期:1977-11-01 00:00:00

  • The molecular basis of vancomycin resistance in clinically relevant Enterococci: crystal structure of D-alanyl-D-lactate ligase (VanA).

    abstract::d-alanine-d-lactate ligase from Enterococcus faecium BM4147 is directly responsible for the biosynthesis of alternate cell-wall precursors in bacteria, which are resistant to the glycopeptide antibiotic vancomycin. The crystal structure has been determined with data extending to 2.5-A resolution. This structure shows ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.150116497

    authors: Roper DI,Huyton T,Vagin A,Dodson G

    更新日期:2000-08-01 00:00:00

  • Evidence for an ancient adaptive episode of convergent molecular evolution.

    abstract::Documented cases of convergent molecular evolution due to selection are fairly unusual, and examples to date have involved only a few amino acid positions. However, because convergence mimics shared ancestry and is not accommodated by current phylogenetic methods, it can strongly mislead phylogenetic inference when it...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0900233106

    authors: Castoe TA,de Koning AP,Kim HM,Gu W,Noonan BP,Naylor G,Jiang ZJ,Parkinson CL,Pollock DD

    更新日期:2009-06-02 00:00:00

  • Sphingosine 1-phosphate receptor modulation suppresses pathogenic astrocyte activation and chronic progressive CNS inflammation.

    abstract::Multiple sclerosis (MS) is an autoimmune inflammatory demyelinating disease of the CNS that causes disability in young adults as a result of the irreversible accumulation of neurological deficits. Although there are potent disease-modifying agents for its initial relapsing-remitting phase, these therapies show limited...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1615413114

    authors: Rothhammer V,Kenison JE,Tjon E,Takenaka MC,de Lima KA,Borucki DM,Chao CC,Wilz A,Blain M,Healy L,Antel J,Quintana FJ

    更新日期:2017-02-21 00:00:00

  • Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells.

    abstract::Although current demands for therapeutic mAbs are growing quickly, production methods to date, including in vitro mammalian tissue culture and transgenic animals, provide only limited quantities at high cost. Several tumor-associated antigens in tumor cells have been identified as targets for therapeutic mAbs. Here we...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0603043103

    authors: Brodzik R,Glogowska M,Bandurska K,Okulicz M,Deka D,Ko K,van der Linden J,Leusen JH,Pogrebnyak N,Golovkin M,Steplewski Z,Koprowski H

    更新日期:2006-06-06 00:00:00

  • Binding of bisubstrate analog promotes large structural changes in the unregulated catalytic trimer of aspartate transcarbamoylase: implications for allosteric regulation.

    abstract::A central problem in understanding enzyme regulation is to define the conformational states that account for allosteric changes in catalytic activity. For Escherichia coli aspartate transcarbamoylase (ATCase; EC) the active, relaxed (R state) holoenzyme is generally assumed to be represented by the crystal structure o...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.090087197

    authors: Endrizzi JA,Beernink PT,Alber T,Schachman HK

    更新日期:2000-05-09 00:00:00

  • Distribution of active and inactive forms of endorphins in rat pituitary and brain.

    abstract::The recent isolation and identification of alpha-N-acetyl forms of the C-Fragment of lipotropin (beta-endorphin, residues 61-91) and the C'-Fragment (residues 61-87) [Smyth, D.G., Massey, D.E., Zakarian, S. & Finnie, M. (1979) Nature (London) 279, 252-254] has led to a study of their distribution in the pituitary and ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.76.11.5972

    authors: Zakarian S,Smyth D

    更新日期:1979-11-01 00:00:00

  • Thermodynamic prediction of protein neutrality.

    abstract::We present a simple theory that uses thermodynamic parameters to predict the probability that a protein retains the wild-type structure after one or more random amino acid substitutions. Our theory predicts that for large numbers of substitutions the probability that a protein retains its structure will decline expone...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0406744102

    authors: Bloom JD,Silberg JJ,Wilke CO,Drummond DA,Adami C,Arnold FH

    更新日期:2005-01-18 00:00:00

  • Characterization of Y73, an avian sarcoma virus: a unique transforming gene and its product, a phosphopolyprotein with protein kinase activity.

    abstract::The Y73 strain of avian sarcoma virus recently isolated in Japan is defective in replication and is associated with subgroup A leukosis virus (YAV). The virus caused sarcoma but not acute leukosis when inoculated into chickens. Studies on the viral RNA showed that a 26S RNA, etimated to be 4.8 kilobases long, was Y73 ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.77.10.6199

    authors: Kawai S,Yoshida M,Segawa K,Sugiyama H,Ishizaki R,Toyoshima K

    更新日期:1980-10-01 00:00:00

  • Histidines, heart of the hydrogen ion channel from influenza A virus: toward an understanding of conductance and proton selectivity.

    abstract::The heart of the H+ conductance mechanism in the homotetrameric M2 H+ channel from influenza A is a set of four histidine side chains. Here, we show that protonation of the third of these imidazoles coincides with acid activation of this transmembrane channel and that, at physiological pH, the channel is closed by two...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0601944103

    authors: Hu J,Fu R,Nishimura K,Zhang L,Zhou HX,Busath DD,Vijayvergiya V,Cross TA

    更新日期:2006-05-02 00:00:00

  • Chromatographic isolation of the hemagglutinin polypeptides from influenza virus vaccine and determination of their amino-terminal sequences.

    abstract::The influenza virus hemagglutinin polypeptides, HA1 and HA2, have been purified by gel filtration in the presence of sodium dodecyl sulfate from a vaccine preparation of the recombinant strain Heq1N2. Use of this technique for purification of the hemagglutinin polypeptides eliminated the need for proteolytic agents fo...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.73.1.238

    authors: Bucher DJ,Li SS,Kehoe JM,Kilbourne ED

    更新日期:1976-01-01 00:00:00

  • Circulating tumor antigen-specific regulatory T cells in patients with metastatic melanoma.

    abstract::Although it is accepted that regulatory T cells (T regs) contribute to cancer progression, most studies in the field consider nonantigen-specific suppression. Here, we show the presence of tumor antigen-specific CD4(+) T regs in the blood of patients with metastatic melanoma. These CD4(+) T regs recognize a broad rang...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0710557105

    authors: Vence L,Palucka AK,Fay JW,Ito T,Liu YJ,Banchereau J,Ueno H

    更新日期:2007-12-26 00:00:00