Plant Cell, Development & Ultrastructure - Botanik in ds9. ?· Plant Cell, Development & Ultrastructure…

  • Published on
    17-Sep-2018

  • View
    212

  • Download
    0

Embed Size (px)

Transcript

  • PCDU Lecture

    izmb.uni-bonn.de izmb.uni-bonn.de

    Plant Cell, Development & Ultrastructure

    Plant Cell Biology Labs

    Download at:

    http://goo.gl/111Tha

  • Endomembranes - Dictyosome / Golgi Apparatus

    Golgi Stack = Dictyosom

    izmb.uni-bonn.de izmb.uni-bonn.de

    Check the ultimate golgi video/song http:/www.brookes.ac.uk/plant_endomembrane

    Golgi Apparatus

  • Dictyosome (Reticulate Body)

    Trans-Side

    Cis-Side

    izmb.uni-bonn.de izmb.uni-bonn.de

    Golgi Apparatus

  • Intercisternal Filaments

    izmb.uni-bonn.de izmb.uni-bonn.de

    100nm

    Hawes 2005

    Tethering Factors

    Barringa et al. 2009

    Golgin

    Giantin

    Golgi Apparatus

  • izmb.uni-bonn.de izmb.uni-bonn.de

    Tethering Factors

    GRIB-domain = C-terminal Arf-binding domain

    named after the first 4 identified members

    GRAB-domain = GRIP-related Arf-binding domain

    GA 1 = GRAB-associated

    Plant Golgin - Candidate Proteins Model of Golgin/G-Protein Interaction

    Arl = Arf-like

    Arf = ADP-ribosylation factor

    Rab = Ras-related in brain

    Ras =rat sarcoma onkogen

    N-Terminus C-Terminus

    Golgi Apparatus

  • GTP Cycle in Small G-Proteins

    GEF = Guanidin Nucleotide Exchange Factor

    GAP = GTPase Activating Protein

    GDI = Guanine Nucleotide Dissociation Inhibitor

    izmb.uni-bonn.de izmb.uni-bonn.de

    Heterotrimeric G-Proteins

  • Retrieval of

    ER-resident Proteins

    Karp 1999

    KDEL Lys Lysin

    Asp Asparagin

    Glu Glutaminsure

    Leu Leucin

    HDEL (Plants)

    izmb.uni-bonn.de izmb.uni-bonn.de

    Endoplasmic Reticulum

  • Retrieval of

    ER-resident Proteins

    Karp 1999

    KDEL Lys Lysin

    Asp Asparagin

    Glu Glutaminsure

    Leu Leucin

    HDEL bei Pflanzen

    izmb.uni-bonn.de izmb.uni-bonn.de

    Regulated by ARF/GEF

    G-Protein

    Endoplasmic Reticulum

  • COPII-Coated Vesicles

    Nickel (1999) Protoplasma 207: 115-124

    ARF/GEF

    GEF = Guanidin Nucleotide Exchange Factor

    BFA

    izmb.uni-bonn.de izmb.uni-bonn.de

    Blickford et al. 2004

    ARF GEFs are Targets of Brefeldin A

  • Fungal Inhibitor Brefeldin A (BFA)

    BFA is a fatty acid metabolite (makrocyclic lactone) from Penicillum brefeldianum. In the

    fungus it is active as an antiviral agent. In animal and plant cells it inhibits anterograde

    vesicle transport from ER to Golgi. Golgi stacks disappear with the consequence that

    exocytosis is abolished. Molecular target is a GTP/GDP-Exchange Factor (GEF) that

    activates small G-proteins involved in the formation of COP-II coated vesicles.

    C16H24O4 MW: 280.36 Da

    1, 13-dihydroxy-6-methyl-4H-cyclopentoxacyclotridecin-4-on

    izmb.uni-bonn.de izmb.uni-bonn.de

    Brefeldin A

  • Effects of BFA on the Endomembrane System

    Nebenfhr (2002) Plant Physiol. 130(3):1102-1108.

    izmb.uni-bonn.de izmb.uni-bonn.de

    Brefeldin A

  • BFA inhibits ER Export

    BFA-Induced Compartments izmb.uni-bonn.de izmb.uni-bonn.de

  • PCR BFA Compartments

    Endocytosis

    Samaj et al., 2004

    BFA inhibits ER Export

    500nm Grebe et al., 2003

    25M BFA, 30min

    izmb.uni-bonn.de izmb.uni-bonn.de

    BFA-Induced Compartments

    12

  • Clathrin-Coated Vesicles for Vesicle Recycling

    36 Triskelions = 12 Pentagons + 6 Hexagons

    izmb.uni-bonn.de izmb.uni-bonn.de

    Clathrin

  • Adaptor Proteins Confer Membrane Domain Specificity

    Receptor-

    Coupled

    Endocytosis

    192 kDa

    25-30 kDa

    izmb.uni-bonn.de izmb.uni-bonn.de

    AP-1 AP-2

    homology

    AP-1

    AP-2

    Clathrin

  • Anti-Transferrin

    15nm gold conjugate

    izmb.uni-bonn.de izmb.uni-bonn.de

    Keyel et al. 2008

    Transferrin is an Fe-transporting

    Glycoprotein that is recognized

    by the transferrin-receptor and

    taken up into the cell by receptor-

    coupled endocytosis

  • Molecular Structure of Clathrin

    izmb.uni-bonn.de izmb.uni-bonn.de

    Clathrin

  • 100kDa Mechanochemical Enzyme:

    Involved in Formation of Coated Vesicles

    Synaptotagmin

    Clathrin

    Amphiphysin Dynamin

    izmb.uni-bonn.de izmb.uni-bonn.de

    Adaptin Complex

    Karp 1999

    Dynamin

  • Bottomley (1999) Curr. Biol. 9: R301-R304

    Recruitment of Dynamin to Membranes

    Pleckstrin-Domain

    PH = Plekstrin-Homology Domain

    GED = GTPase regulatory Domain

    PRD = Prolin rich Domain

    izmb.uni-bonn.de izmb.uni-bonn.de

    Dynamin

  • Intracellular Motility of Golgi Stacks in Animals Cells

    is Dependent on the Microtubule Cytoskeleton

    izmb.uni-bonn.de izmb.uni-bonn.de

    Dictyosomes Dictyosomes

    plus actin

    Microtubules in

    mouse fibroblasts, Jan Schmoranzer

    http://www.maths.bris.ac.uk

    Golgi in HeLa cell

    Abcam plc 2008

    Golgi Stacks in Animal Cells

  • Intracellular Motility of Golgi Stacks (Dictyosomes) in

    Plant Cells is Dependent on the Actin Cytoskeleton

    izmb.uni-bonn.de izmb.uni-bonn.de

    Dictyosomes Dictyosomes

    plus actin

    ER

    Chris Hawes

    Golgi Stacks

    (Dictyosomes)

    Golgi Stacks in Plant Cells

  • Secretory Pathway to Vacuoles

    izmb.uni-bonn.de izmb.uni-bonn.de

  • Prevacuolar Compartment

    PVC

    izmb.uni-bonn.de izmb.uni-bonn.de

    ESCRT = Endosomal Sorting Complex Required for Transport

    MVB = Multi-Vesicular Body

    Retromer = Rretrograde Transport Complex to the TGN

    Otegui & Spitzer 2008

    Prevacuolar Compartment PVC

    Multivesicular Body MVB

    ESCRT Complex - ESCRT

  • Retromer Complex izmb.uni-bonn.de izmb.uni-bonn.de

    Retromer = Pentameric Protein Complex for the Retrieval of VSR

    VSR = Vacuolar Sorting Receptor

    26/29/35 = VPS proteins (Vacuolar Protein Sorting Proteins)

    SNX = Sorting Nexin

    PI3P = Phosphatidylinositol 3-Phosphate

    coiled-coil

    PI-3-P-binding

    Transport of Cargo from MVB Back to TGN

    Schellmann & Pimpl 2009

  • Dynamics within Golgi Stack:

    Stationary Cisternae Model

    versus Vesicle Shuttle Model

    Buchanan et al. 2000

    Intercisternal

    elements

    izmb.uni-bonn.de izmb.uni-bonn.de

    Dynamics within Golgi Stack

  • izmb.uni-bonn.de izmb.uni-bonn.de

    Dynamics within Golgi Stack

  • Dictyosomes in Coccolith Algae

    Support the Cisternal Progression Model

    Cross-sectional model of coccolith formation

    in the Golgi of P. carterae.

    (1) Polysaccharide scale formation

    (2) Mineral deposition

    (3) Mature Coccosphere

    P = Ca2+-containing particles

    Coccolithophora

    izmb.uni-bonn.de izmb.uni-bonn.de

    Dynamics within Golgi Stack

  • Synthetic Functions of Golgi Stacks

    Hemicelluloses Pectins

    Aktivierte Monosaccharide

    Transporter

    izmb.uni-bonn.de izmb.uni-bonn.de

  • ATP

    ADP

    Uncoating ATPases

    NSF= Nethylmaleimid-Sensitive Factor (triple helix resolving ATPase)

    SNAP= soluble N-ethylmaleimide-sensitive-factor Attachment Protein

    SNARE= soluble N-ethylmaleimide-sensitive-factor Attachment-Protein REceptor

    NSF

    izmb.uni-bonn.de izmb.uni-bonn.de

  • The SNARE Hypothesis

    Explains Targeted Vesicle Fusion

    izmb.uni-bonn.de izmb.uni-bonn.de

    SNARE Hypothesis

    Exocyst

  • (v-snare)

    (v-snare)

    (t-snare) tethering

    complex

    Kiss & Run

    izmb.uni-bonn.de izmb.uni-bonn.de

    Vesicle Fusion with Plasma Membrane

  • izmb.uni-bonn.de izmb.uni-bonn.de

    Bai & Chapman 2004

    SYT1 Controls Membrane Repair in Plant Cells

    Synaptotagmin

    Synaptotagmins in Plants

  • Microbodies/ Peroxisomes

    izmb.uni-bonn.de izmb.uni-bonn.de

    1

  • Source for H2O2 in Catabolic pathways

    izmb.uni-bonn.de izmb.uni-bonn.de

    Katalase

    Uricosomen in Stickstoff-speichernden

    Geweben

    Glyoxysomen in Stickstoff-speichernden

    Geweben

    Peroxisomen in Blattgewebe

    2

  • Nitrogen storage in uricosomes

    izmb.uni-bonn.de izmb.uni-bonn.de

    3

  • Histochemical test for catalase

    izmb.uni-bonn.de izmb.uni-bonn.de

    DAB

    H2O2

    + DAB

    Diaminobenzidin

    4

  • izmb.uni-bonn.de izmb.uni-bonn.de

    Import of peroxisomal membrane proteins

    Peroxisomal membrane proteins (PMPs) enter the ER, are sequestered in specialized regions and bud from the ER to form peroxisomal pre-

    compartments. These become capable of importing PTS1/PTS2 containing proteins and develop into mature peroxisomes or fuse with existing

    peroxisomes. Enlarged peroxisomes can divide by fission.

    Lipid import

    mature peroxisome pre-peroxisome

    5

  • izmb.uni-bonn.de izmb.uni-bonn.de

    Import of peroxisomal membrane proteins

    NBF = Nucleotide binding fold

    TMD = Transmembrane domain

    ABC = A