INTRAFLAGELLAR TRANSPORT IN TRYPANOSOMES Filippo RUSCONI Muséum National d’Histoire Naturelle INSERM U565 & CNRS UMR 5153 & MNHN 0503

Slides:

Advertisements

Similar presentations

Problem Results: Question: 1. You screen two libraries- cDNA; genomic

Advertisements

CELL STRUCTURE & FUNCTION

Lecture 19 Homework Review Today: Apoptosis and Cancer Monday April 23rd- Cell-Cell Interactions/Tissues Thursday April 26 th -Early Development and Stem.

Cell identity and positional information. How does a neuron find its target?

Cell and Molecular Biology Behrouz Mahmoudi Cell cycle 1.

Molecular Motors Filament…….motor Actin…….….myosins Tubulin….….dynein and kinesins.

The Cytoskeleton. Intermediate Filaments 8-10 nm in diameter Four types: basis of aa sequence actin & tubulin -> globular protein, cell type.

Introduction to yeast genetics Michelle Attner July 24, 2012.

Work Process Using Enrich Load biological data Check enrichment of crossed data sets Extract statistically significant results Multiple hypothesis correction.

©CMBI 2006 Molecular motors At the organism – population level, motors are needed to transfer materials. At the organelle – organ level, motors are needed.

Lecture 11 Vesicular Trafficking: Lysosomes;

Cell Motility Lecture 17. Cell Motility Includes: –Changes in Cell Location –Limited Movements of Parts of Cells Occurs at the Subcellular, Cellular,

CDC42 a low molecular weight GTP-binding protein originally designated G(p) and also called G25K Chromosomal location: 1p36.1 The CDC42 protein binds to.

Investigating the Role of Inner-arm Dynein Knockdowns in Trypanosoma brucei Kathryn Kinzel.

Discovery of Alternative Splicing

Genetics Cell Cycle Mitosis Meiosis.

Microtubules (17) Dynamic instability –Growing and shrinking microtubules can coexist in the same region of a cell. –A given microtubule can switch back.

Cell Introduction Prokaryotic Cell: A cell that is lacking a nucleus and most organelles Prokaryotic Cell: A cell that is lacking a nucleus and most organelles.

Basic DNA Science BE Bootcamp 2008 Phillips Group / Caltech.

The cytoskeleton, mitochondrial bioenergetics and apoptosis Professor Daniel C. Hoessli March 2013.

F LAGELLUM /C ILIUM A Presentation By: Katie Scheuch.

rasiRNAs: a class of small regulatory RNAs

Pages Molecular Motors. General Characteristics of Molecular Motors Motor proteins – bind to a polarized cytoskeletal filament and use the energy.

The Genetic Basis of Development

CHAPTER 9 The Cytoskeleton and Cell Motility. Introduction The cytoskeleton is a network of filamentous structures: microtubulues, microfilaments, and.

AP Biology Tour of the Cell 3 AP Biology Cells gotta work to live!  What jobs do cells have to do?  make proteins  proteins control every.

Centrioles. The Centrosome and the Centrioles ANIMAL CELL CENTROSOME: The centrosome, also called the "microtubule organizing center", is an area in the.

Cell Life Cycle. Cells have two major periods Interphase Cell grows Cell carries on metabolic processes Cell replicates DNA Cell division Cell replicates.

Filaments Of The Cytoskeleton

CELL DIVISION. Stages of mitosis (animal cell) prophase: - chromosomes condense (replicated in S phase) - centrosomes separate (duplicated in S phase)

Javad Jamshidi Fasa University of Medical Sciences, October 2015 Eukaryotic Cell Organelles and Organization.

BIOLOGY OF HUMAN AGING CHAPTER 3 CELLULAR AGING. Cell components * Organelles * Nucleus * Cytoplasm * Plasma membrane.

Fundaments of Cell Biology Kristen Lee 9/23/2009.

RanBP3 enhances nuclear export of active β-catenin independently of CRM1 Hendriksen et al., JCB(2005)

Intro to topic 3 Cell Division – one cell divides to make 2 new daughter cells.

Zebrafish in genome research

Cytoskeleton and Cell Motility. Objective To survey the structure, protein composition, and functions of a complex network of cytoplasmic filaments known.

Cell Cycle Biology I. Cell Cycle = sequence of growth and division in the life of a cell = sequence of growth and division in the life of a cell Animation.

AP Biology Eukaryotic Genome Control Mechanisms for Gene expression.

Vectors for RNAi.

Knowledge prior to this research? Questions addressed? Experimental approaches used? and what outcome? Impact of these findings? Future experiments? Research.

Excellent cell biology hormone and light signaling pathways operative few cell types & simple developmental pattern polar tip growth and division tolerant.

The Cytoskeleton Functions

Cytoskeleton Chapter Pgs Objective: I can describe how the cytoskeleton is structured based on different protein fibers, and based on these,

The Moving Parts: Microtubules and MT- based Motors. MULTIGENE FAMILIES: C. elegans MTs: 9 α–tubulins 6 β-tubulins 1 γ-tubulin C. elegans MT-motors: -21.

7.3 Translation udent_view0/chapter3/animation__how_translation_work s.html.

The cytoskeleton: (Fig.4.19 page 76)

Vesicular transport Dr. med. habil. Kőhidai László Assoc. Professor

Organelle Transport: A Park-and-Ride System for Melanosomes

Lotte B. Pedersen, Stefan Geimer, Joel L. Rosenbaum Current Biology

HIS-24 regulates expression of infection-inducible genes.

Jack D. Sunter, Keith Gull Trends in Parasitology

General Animal Biology

Inferring chromatin organization.

Chlamydomonas Shortens Its Flagella by Activating Axonemal Disassembly, Stimulating IFT Particle Trafficking, and Blocking Anterograde Cargo Loading

Volume 30, Issue 5, Pages (September 2014)

Cilia in the CNS: The Quiet Organelle Claims Center Stage

Nat. Rev. Neurol. doi: /nrneurol

Tomer Avidor-Reiss, Michel R. Leroux Current Biology

Membrane Traffic in the Late Steps of Cytokinesis

RND efflux operons in P. aeruginosa.

Volume 3, Issue 5, Pages (November 2002)

Marco Terenzio, Giampietro Schiavo, Mike Fainzilber Neuron

Volume 19, Issue 20, Pages (November 2009)

Volume 11, Issue 20, Pages (October 2001)

Fig. 6. Hts regulates par-1 and camkII mRNA distribution and levels in the muscle.(A–J) Views of muscles 6/7 in abdominal segment 4, probed for either.

It is composed of three types of molecular structures:

Drug transport mechanism of AcrB.

Volume 14, Issue 14, Pages R575-R577 (July 2004)

Presentation transcript:

INTRAFLAGELLAR TRANSPORT IN TRYPANOSOMES Filippo RUSCONI Muséum National d’Histoire Naturelle INSERM U565 & CNRS UMR 5153 & MNHN 0503

IntraFlagellar Transport (IFT) Radial spoke Anterograde IFT : FLA10 Kinesin II (towards flagellum tip, + end MT) Retrograde IFT: Cytoplasmic Dynein DHC1b (towards cell body) IFT particle Raft FLA10 KinesinII Cytoplasmic Dynein Cole et al., J Cell Biol. 141, , 1998

PhaseAnti-flagellum + DAPI nucleus Trypanosoma brucei (protozoan) Flagellated organism easy to grow in vitro Genome sequenced Straight forward molecular biology Reverse genetics (RNAi, inducible expression) Responsible for sleeping sickness Flagellum attached to the cell body

IFT in trypanosomes PFRA T7 tet-inducible promoters T7 transcription terminators drug resistance Wang et al., JBC (2000) 275, Durand-Dubief, Kohl & Bastin, MBP (2003) 129, intermolecular GeneCharacteristicsIFT in T. b. Motors DHC1bdynein heavy chain+ DHC1bisdynein heavy chain+ LC8dynein light chain- KIF3Akinesin motor- KIF3Bkinesin motorND KAPcargo bindingND Raft components IFT20coiled-coil+ IFT27G protein? IFT52coiled-coil+ IFT55coiled-coil+ IFT74coiled-coilND IFT88TPR+ IFT122WD40+ IFT172WD40+ Regulators LF4kinase+/- Vector for RNAi silencing

The trypanosome cell cycle Green: Flagellum, Red lines: FAZ, Red dots: basal body, Blue: DAPI

Generation of non-flagellated cells non- induced Green: Flagellum, Red lines: FAZ, Red dots: basal body, Blue: DAPI induced Defects:No new flagellum, but FAZ is still there but... FAZ is too short

Shorter flagella, shorter cells ? Green: Flagellum, Red lines: FAZ, Red dots: basal body, Blue: DAPI DIC (TbDHC1b cells, RNAi silencing induced for 48 hrs)

Flagellum length (µm) cell size (µm) Flagellum and cell size are linked

DHC1b IFT88 Induced cells don’t have a flagellum Characterisation of IFT RNAi mutants % Flagellated cells

FAZ FORMATION AND CYTOKINESIS no new flagellum new FAZ no new flagellum no new FAZ no new flagellum no new FAZ No cytokinesis Nuclear division OK Blue: DAPI Red lines: FAZ Red spots: basal bodies Non- flagellated sibling Flagellated sibling Multinucleated cells

Summary 1) IFT controls flagellum assembly 2) IFT controls flagellum length 3) Flagellum length dictates FAZ elongation and defines cytokinesis initiation 4) Flagellum and FAZ are required for cell morphogenesis (shape, polarity) Kohl, Robinson & Bastin, EMBO J. (2003) 22, See also comments in Hurtley S. (2003) A dead-end short cut. Science 302, 535 Jones S. (2003) Trypanosome flagellum-motility and more. Nature Rev. Microbiol. 1,

Non-flagellated cells show dispersion of their endocytic network Red: anti-clathrin Normal Non-flagellated Blue: DAPI Dispersion of : -rab5 (early endosomes) -rab11 (recycling pathway) -p67 (lysosomes)

Group Philippe BASTIN (Muséum National d’Histoire Naturelle, Paris, Dir. C. GIOVANNANGELI & J.S. SUN) Linda KOHL Filippo RUSCONI Sabrina BENGHANEM Géraldine TOUTIRAIS Mickaël DURAND-DUBIEF Carole BRANCHE Gwénola DORE Derrick ROBINSON (Université Bordeaux II) Mark FIELD (Imperial College, UK) Thanks to K. GULL, P. ENGLUND, C. CLAYTON, G. CROSS