Genetically inspired in vitro reconstitution of Saccharomyces cerevisiae actin cables from seven purified proteins.

Abstract:

:A major goal of synthetic biology is to define the minimal cellular machinery required to assemble a biological structure in its simplest form. Here, we focused on Saccharomyces cerevisiae actin cables, which provide polarized tracks for intracellular transport and maintain defined lengths while continuously undergoing rapid assembly and turnover. Guided by the genetic requirements for proper cable assembly and dynamics, we show that seven evolutionarily conserved S. cerevisiae proteins (actin, formin, profilin, tropomyosin, capping protein, cofilin, and AIP1) are sufficient to reconstitute the formation of cables that undergo polarized turnover and maintain steady-state lengths similar to actin cables in vivo. Further, the removal of individual proteins from this simple in vitro reconstitution system leads to cable defects that closely approximate in vivo cable phenotypes caused by disrupting the corresponding genes. Thus, a limited set of molecular components is capable of self-organizing into dynamic, micron-scale actin structures with features similar to cables in living cells.

journal_name

Mol Biol Cell

authors

Pollard LW,Garabedian MV,Alioto SL,Shekhar S,Goode BL

doi

10.1091/mbc.E19-10-0576

subject

Has Abstract

pub_date

2020-03-01 00:00:00

pages

335-347

issue

5

eissn

1059-1524

issn

1939-4586

journal_volume

31

pub_type

杂志文章
  • Mutations in a beta-tubulin disrupt spindle orientation and microtubule dynamics in the early Caenorhabditis elegans embryo.

    abstract::The early Caenorhabditis elegans embryo contains abundant transcripts for two alpha- and two beta-tubulins, raising the question of whether each isoform performs specialized functions or simply contributes to total tubulin levels. Our identification of two recessive, complementing alleles of a beta-tubulin that disrup...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e03-01-0017

    authors: Wright AJ,Hunter CP

    更新日期:2003-11-01 00:00:00

  • Functional analysis of the interaction between Afr1p and the Cdc12p septin, two proteins involved in pheromone-induced morphogenesis.

    abstract::Saccharomyces cerevisiae mating pheromones induce production of Afr1p, a protein that negatively regulates pheromone receptor signaling and is required for normal formation of the projection of cell growth that becomes the site of cell fusion during conjugation. Afr1p interacts with Cdc12p, which belongs to a family o...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.8.6.987

    authors: Giot L,Konopka JB

    更新日期:1997-06-01 00:00:00

  • Histone deacetylase inhibitors trigger a G2 checkpoint in normal cells that is defective in tumor cells.

    abstract::Important aspects of cell cycle regulation are the checkpoints, which respond to a variety of cellular stresses to inhibit cell cycle progression and act as protective mechanisms to ensure genomic integrity. An increasing number of tumor suppressors are being demonstrated to have roles in checkpoint mechanisms, implyi...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.11.6.2069

    authors: Qiu L,Burgess A,Fairlie DP,Leonard H,Parsons PG,Gabrielli BG

    更新日期:2000-06-01 00:00:00

  • Folding of active beta-lactamase in the yeast cytoplasm before translocation into the endoplasmic reticulum.

    abstract::Polypeptides targeted to the yeast endoplasmic reticulum (ER) posttranslationally are thought to be kept in the cytoplasm in an unfolded state by Hsp70 chaperones before translocation. We show here that Escherichia coli beta-lactamase associated with Hsp70, but adopted a native-like conformation before translocation i...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.9.4.817

    authors: Paunola E,Suntio T,Jämsä E,Makarow M

    更新日期:1998-04-01 00:00:00

  • Protein targeting of an unusual, evolutionarily conserved adenylate kinase to a eukaryotic flagellum.

    abstract::The eukaryotic flagellum is a large structure into which specific constituent proteins must be targeted, transported and assembled after their synthesis in the cytoplasm. Using Trypanosoma brucei and a proteomic approach, we have identified and characterized a novel set of adenylate kinase proteins that are localized ...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e04-03-0217

    authors: Pullen TJ,Ginger ML,Gaskell SJ,Gull K

    更新日期:2004-07-01 00:00:00

  • The role actin filaments play in providing the characteristic curved form of Drosophila bristles.

    abstract::Drosophila bristles display a precise orientation and curvature. An asymmetric extension of the socket cell overlies the newly emerging bristle rudiment to provide direction for bristle elongation, a process thought to be orchestrated by the nerve dendrite lying between these cells. Scanning electron microscopic analy...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e04-06-0472

    authors: Tilney LG,Connelly PS,Ruggiero L,Vranich KA,Guild GM,Derosier D

    更新日期:2004-12-01 00:00:00

  • Functional analysis and importance for host cell infection of the Ca2+-conducting subunits of the mitochondrial calcium uniporter of Trypanosoma cruzi.

    abstract::We report here that Trypanosoma cruzi, the etiologic agent of Chagas disease, possesses two unique paralogues of the mitochondrial calcium uniporter complex TcMCU subunit that we named TcMCUc and TcMCUd. The predicted structure of the proteins indicates that, as predicted for the TcMCU and TcMCUb paralogues, they are ...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E19-03-0152

    authors: Chiurillo MA,Lander N,Bertolini MS,Vercesi AE,Docampo R

    更新日期:2019-07-01 00:00:00

  • Hydrogen sulfide increases hypoxia-inducible factor-1 activity independently of von Hippel-Lindau tumor suppressor-1 in C. elegans.

    abstract::Rapid alteration of gene expression in response to environmental changes is essential for normal development and behavior. The transcription factor hypoxia-inducible factor (HIF)-1 is well known to respond to alterations in oxygen availability. In nature, low oxygen environments are often found to contain high levels ...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e09-03-0199

    authors: Budde MW,Roth MB

    更新日期:2010-01-01 00:00:00

  • Conserved Ark1-related kinases function in a TORC2 signaling network.

    abstract::In all orders of life, cell cycle progression in proliferating cells is dependent on cell growth, and the extent of growth required for cell cycle progression is proportional to growth rate. Thus, cells growing rapidly in rich nutrients are substantially larger than slow-growing cells. In budding yeast, a conserved si...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E19-12-0685

    authors: Alcaide-Gavilán M,Lucena R,Banuelos S,Kellogg DR

    更新日期:2020-08-15 00:00:00

  • Glycoprotein gp130 of dictyostelium discoideum influences macropinocytosis and adhesion.

    abstract::Glycoprotein gp130, found on the plasma membrane of Dictyostelium discoideum amoebae, was postulated previously to play a role in phagocytosis. The gene for gp130 was cloned and when translated, yielded a 768 amino acid preproprotein of 85.3 kDa. It had nearly 40% similarity to the 138 kDa family of glycoproteins impl...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e04-06-0483

    authors: Chia CP,Gomathinayagam S,Schmaltz RJ,Smoyer LK

    更新日期:2005-06-01 00:00:00

  • ATP-2 interacts with the PLAT domain of LOV-1 and is involved in Caenorhabditis elegans polycystin signaling.

    abstract::Caenorhabditis elegans is a powerful model to study the molecular basis of autosomal dominant polycystic kidney disease (ADPKD). ADPKD is caused by mutations in the polycystic kidney disease (PKD)1 or PKD2 gene, encoding polycystin (PC)-1 or PC-2, respectively. The C. elegans polycystins LOV-1 and PKD-2 are required f...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e04-09-0851

    authors: Hu J,Barr MM

    更新日期:2005-02-01 00:00:00

  • Energy metabolism regulates clathrin adaptors at the trans-Golgi network and endosomes.

    abstract::Glucose is a master regulator of cell behavior in the yeast Saccharomyces cerevisiae. It acts as both a metabolic substrate and a potent regulator of intracellular signaling cascades. Glucose starvation induces the transient delocalization and then partial relocalization of clathrin adaptors at the trans-Golgi network...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E12-10-0750

    authors: Aoh QL,Hung CW,Duncan MC

    更新日期:2013-03-01 00:00:00

  • The 90-kDa heat shock protein stabilizes the polysomal ribonuclease 1 mRNA endonuclease to degradation by the 26S proteasome.

    abstract::The polysomal ribonuclease 1 (PMR1) mRNA endonuclease forms a selective complex with its translating substrate mRNAs where it is activated to initiate mRNA decay. Previous work showed tyrosine phosphorylation is required for PMR1 targeting to this polysome-bound complex, and it identified c-Src as the responsible kina...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e07-08-0774

    authors: Peng Y,Liu X,Schoenberg DR

    更新日期:2008-02-01 00:00:00

  • Fission yeast sts1+ gene encodes a protein similar to the chicken lamin B receptor and is implicated in pleiotropic drug-sensitivity, divalent cation-sensitivity, and osmoregulation.

    abstract::The Schizosaccharomyces pombe sts1+ gene, identified by supersensitive mutations to a protein kinase inhibitor, staurosporine, was isolated by complementation by the use of a fission yeast genomic library. Nucleotide sequencing shows that the sts1+ gene encodes a 453 amino acid putative membrane-associated protein tha...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.3.3.263

    authors: Shimanuki M,Goebl M,Yanagida M,Toda T

    更新日期:1992-03-01 00:00:00

  • The role of HYAL2 in LSS-induced glycocalyx impairment and the PKA-mediated decrease in eNOS-Ser-633 phosphorylation and nitric oxide production.

    abstract::Hyaluronan (HA) in the endothelial glycocalyx serves as a mechanotransducer for high-shear-stress-stimulated endothelial nitric oxide synthase (eNOS) phosphorylation and nitric oxide (NO) production. Low shear stress (LSS) has been shown to contribute to endothelial inflammation and atherosclerosis by impairing the ba...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E16-04-0241

    authors: Kong X,Chen L,Ye P,Wang Z,Zhang J,Ye F,Chen S

    更新日期:2016-12-15 00:00:00

  • The HOPS complex mediates autophagosome-lysosome fusion through interaction with syntaxin 17.

    abstract::Membrane fusion is generally controlled by Rabs, soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), and tethering complexes. Syntaxin 17 (STX17) was recently identified as the autophagosomal SNARE required for autophagosome-lysosome fusion in mammals and Drosophila. In this study, to bett...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E13-08-0447

    authors: Jiang P,Nishimura T,Sakamaki Y,Itakura E,Hatta T,Natsume T,Mizushima N

    更新日期:2014-04-01 00:00:00

  • Cysteine string protein functions directly in regulated exocytosis.

    abstract::Cysteine string protein (Csp) is essential for neurotransmitter release in Drosophila. It has been suggested that Csp functions by regulating the activity of presynaptic Ca2+ channels, thus controlling exocytosis. We have examined the effect of overexpressing Csp1 in PC12 cells, a neuroendocrine cell line. PC12 cell c...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.9.8.2259

    authors: Chamberlain LH,Burgoyne RD

    更新日期:1998-08-01 00:00:00

  • Impaired fertility and spermiogenetic disorders with loss of cell adhesion in male mice expressing an interfering Rap1 mutant.

    abstract::The guanosine trisphosphatase Rap1 serves as a critical player in signal transduction, somatic cell proliferation and differentiation, and cell-cell adhesion by acting through distinct mechanisms. During mouse spermiogenesis, Rap1 is activated and forms a signaling complex with its effector, the serine-threonine kinas...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e06-10-0902

    authors: Aivatiadou E,Mattei E,Ceriani M,Tilia L,Berruti G

    更新日期:2007-04-01 00:00:00

  • Plc1p is required for SAGA recruitment and derepression of Sko1p-regulated genes.

    abstract::In Saccharomyces cerevisiae, many osmotically inducible genes are regulated by the Sko1p-Ssn6p-Tup1p complex. On osmotic shock, the MAP kinase Hog1p associates with this complex, phosphorylates Sko1p, and converts it into an activator that subsequently recruits Swi/Snf and SAGA complexes. We have found that phospholip...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e06-10-0946

    authors: Guha N,Desai P,Vancura A

    更新日期:2007-07-01 00:00:00

  • Phosphorylation of a conserved integrin alpha 3 QPSXXE motif regulates signaling, motility, and cytoskeletal engagement.

    abstract::Integrin alpha 3A cytoplasmic tail phosphorylation was mapped to amino acid S1042, as determined by mass spectrometry, and confirmed by mutagenesis. This residue occurs within a "QPSXXE" motif conserved in multiple alpha chains (alpha 3A, alpha 6A, alpha 7A), from multiple species. Phosphorylation of alpha 3A and alph...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.12.2.351

    authors: Zhang XA,Bontrager AL,Stipp CS,Kraeft SK,Bazzoni G,Chen LB,Hemler ME

    更新日期:2001-02-01 00:00:00

  • Agonist-induced PIP(2) hydrolysis inhibits cortical actin dynamics: regulation at a global but not at a micrometer scale.

    abstract::Phosphatidylinositol 4, 5-bisphosphate (PIP(2)) at the inner leaflet of the plasma membrane has been proposed to locally regulate the actin cytoskeleton. Indeed, recent studies that use GFP-tagged pleckstrin homology domains (GFP-PH) as fluorescent PIP(2) sensors suggest that this lipid is enriched in membrane microdo...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e02-04-0231

    authors: van Rheenen J,Jalink K

    更新日期:2002-09-01 00:00:00

  • Prostaglandins temporally regulate cytoplasmic actin bundle formation during Drosophila oogenesis.

    abstract::Prostaglandins (PGs)--lipid signals produced downstream of cyclooxygenase (COX) enzymes--regulate actin dynamics in cell culture and platelets, but their roles during development are largely unknown. Here we define a new role for Pxt, the Drosophila COX-like enzyme, in regulating the actin cytoskeleton--temporal restr...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E13-07-0366

    authors: Spracklen AJ,Kelpsch DJ,Chen X,Spracklen CN,Tootle TL

    更新日期:2014-02-01 00:00:00

  • The stem-loop binding protein (SLBP1) is present in coiled bodies of the Xenopus germinal vesicle.

    abstract::The stem-loop binding protein (SLBP1) binds the 3' stem-loop of histone pre-mRNA and is required for efficient processing of histone transcripts in the nucleus. We examined the localization of SLBP1 in the germinal vesicle of Xenopus laevis oocytes. In spread preparations of germinal vesicle contents, an anti-SLBP1 an...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.10.2.487

    authors: Abbott J,Marzluff WF,Gall JG

    更新日期:1999-02-01 00:00:00

  • Mrx6 regulates mitochondrial DNA copy number in Saccharomyces cerevisiae by engaging the evolutionarily conserved Lon protease Pim1.

    abstract::Mitochondrial function depends crucially on the maintenance of multiple mitochondrial DNA (mtDNA) copies. Surprisingly, the cellular mechanisms regulating mtDNA copy number remain poorly understood. Through a systematic high-throughput approach in Saccharomyces cerevisiae, we determined mtDNA-to-nuclear DNA ratios in ...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E19-08-0470

    authors: Göke A,Schrott S,Mizrak A,Belyy V,Osman C,Walter P

    更新日期:2020-03-19 00:00:00

  • Neurodegeneration-associated mutant TREM2 proteins abortively cycle between the ER and ER-Golgi intermediate compartment.

    abstract::Triggering receptor expressed on myeloid cells 2 (TREM2) is a transmembrane protein expressed on microglia within the brain. Several rare mutations in TREM2 cause an early-onset form of neurodegeneration when inherited homozygously. Here we investigate how these mutations affect the intracellular transport of TREM2. W...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E17-06-0423

    authors: Sirkis DW,Aparicio RE,Schekman R

    更新日期:2017-10-01 00:00:00

  • Iterative sorting of apical and basolateral cargo in Madin-Darby canine kidney cells.

    abstract::For several decades, the trans-Golgi network (TGN) was considered the most distal stop and hence the ultimate protein-sorting station for distinct apical and basolateral transport carriers that reach their respective surface domains in the direct trafficking pathway. However, recent reports of apical and basolateral c...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E16-02-0096

    authors: Treyer A,Pujato M,Pechuan X,Müsch A

    更新日期:2016-07-15 00:00:00

  • Paradoxical instability-activity relationship defines a novel regulatory pathway for retinoblastoma proteins.

    abstract::The retinoblastoma (RB) transcriptional corepressor and related family of pocket proteins play central roles in cell cycle control and development, and the regulatory networks governed by these factors are frequently inactivated during tumorigenesis. During normal growth, these proteins are subject to tight control th...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.E10-06-0520

    authors: Acharya P,Raj N,Buckley MS,Zhang L,Duperon S,Williams G,Henry RW,Arnosti DN

    更新日期:2010-11-15 00:00:00

  • Role for actin filament turnover and a myosin II motor in cytoskeleton-driven disassembly of the epithelial apical junctional complex.

    abstract::Disassembly of the epithelial apical junctional complex (AJC), composed of the tight junction (TJ) and adherens junction (AJ), is important for normal tissue remodeling and pathogen-induced disruption of epithelial barriers. Using a calcium depletion model in T84 epithelial cells, we previously found that disassembly ...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e04-02-0163

    authors: Ivanov AI,McCall IC,Parkos CA,Nusrat A

    更新日期:2004-06-01 00:00:00

  • Reorganization of actin cytoskeleton by the phosphoinositide metabolite glycerophosphoinositol 4-phosphate.

    abstract::Glycerophosphoinositol 4-phosphate (GroPIns-4P) is a biologically active, water-soluble phospholipase A metabolite derived from phosphatidylinositol 4-phosphate, whose cellular concentrations have been reported to increase in Ras-transformed cells. It is therefore important to understand its biological activities. Her...

    journal_title:Molecular biology of the cell

    pub_type: 杂志文章

    doi:10.1091/mbc.e02-04-0179

    authors: Mancini R,Piccolo E,Mariggio' S,Filippi BM,Iurisci C,Pertile P,Berrie CP,Corda D

    更新日期:2003-02-01 00:00:00

  • Morphogenesis in Kyoto: a confluence of cell and developmental biology.

    abstract::Understanding morphogenesis is the ultimate multidisciplinary (ad)venture. Three-dimensional tissues are generated from the actions of genes, biochemical pathways, and cells that form multicellular networks and interact with their biomechanical environment. A comprehensive explanation of morphogenetic processes must e...

    journal_title:Molecular biology of the cell

    pub_type:

    doi:10.1091/mbc.e09-12-1076

    authors: Zallen JA,Yap AS

    更新日期:2010-03-15 00:00:00