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PMID: 23918939 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Lissencephaly-1 promotes the recruitment of dynein and dynactin to transported mRNAs.

The Journal of cell biology ·Vol. 202 ·No. 3 ·2013-08-05 ·Pages 479-94

Dix CI, Soundararajan HC, Dzhindzhev NS, Begum F, Suter B, Ohkura H, Stephens E, Bullock SL

Abstract

Microtubule-based transport mediates the sorting and dispersal of many cellular components and pathogens. However, the mechanisms by which motor complexes are recruited to and regulated on different cargos remain poorly understood. Here we describe a large-scale biochemical screen for novel factors associated with RNA localization signals mediating minus end-directed mRNA transport during Drosophila development. We identified the protein Lissencephaly-1 (Lis1) and found that minus-end travel distances of localizing transcripts are dramatically reduced in lis1 mutant embryos. Surprisingly, given its well-documented role in regulating dynein mechanochemistry, we uncovered an important requirement for Lis1 in promoting the recruitment of dynein and its accessory complex dynactin to RNA localization complexes. Furthermore, we provide evidence that Lis1 levels regulate the overall association of dynein with dynactin. Our data therefore reveal a critical role for Lis1 within the mRNA localization machinery and suggest a model in which Lis1 facilitates motor complex association with cargos by promoting the interaction of dynein with dynactin.

MeSH Terms
Animals Drosophila/metabolism Drosophila Proteins/metabolism Dynactin Complex Dyneins/metabolism Microtubule-Associated Proteins/metabolism Protein Transport RNA Transport RNA, Messenger/metabolism
Chemicals
Drosophila Proteins Dynactin Complex Lis-1 protein, Drosophila Microtubule-Associated Proteins RNA, Messenger Dyneins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Dix Carly I
Cell Biology Division, MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, England, UK.
Soundararajan Harish Chandra
Dzhindzhev Nikola S
Begum Farida
Suter Beat
Ohkura Hiroyuki
Stephens Elaine
Bullock Simon L
References (86)
86 references, click to expand
  1. Processive bidirectional motion of dynein-dynactin complexes in vitro.
    Nat Cell Biol. 2006 Jun;8(6):562-70 PMID: 16715075
  2. Dynactin suppresses the retrograde movement of apically localized mRNA in Drosophila blastoderm embryos.
    RNA. 2007 Nov;13(11):1860-7 PMID: 17901156
  3. The involvement of the intermediate chain of cytoplasmic dynein in binding the motor complex to membranous organelles of Xenopus oocytes.
    Mol Biol Cell. 1997 Oct;8(10):2077-88 PMID: 9348543
  4. Neuroepithelial stem cell proliferation requires LIS1 for precise spindle orientation and symmetric division.
    Cell. 2008 Feb 8;132(3):474-86 PMID: 18267077
  5. The Drosophila hairy RNA localization signal modulates the kinetics of cytoplasmic mRNA transport.
    EMBO J. 2003 May 15;22(10):2484-94 PMID: 12743042
  6. Distinct mechanisms govern the localisation of Drosophila CLIP-190 to unattached kinetochores and microtubule plus-ends.
    J Cell Sci. 2005 Aug 15;118(Pt 16):3781-90 PMID: 16105886
  7. Drosophila wingless and pair-rule transcripts localize apically by dynein-mediated transport of RNA particles.
    Cell. 2001 Apr 20;105(2):209-19 PMID: 11336671
  8. lis-1 is required for dynein-dependent cell division processes in C. elegans embryos.
    J Cell Sci. 2004 Sep 1;117(Pt 19):4571-82 PMID: 15331665
  9. A role for the lissencephaly gene LIS1 in mitosis and cytoplasmic dynein function.
    Nat Cell Biol. 2000 Nov;2(11):784-91 PMID: 11056532
  10. Graded reduction of Pafah1b1 (Lis1) activity results in neuronal migration defects and early embryonic lethality.
    Nat Genet. 1998 Aug;19(4):333-9 PMID: 9697693
  11. Consequences of motor copy number on the intracellular transport of kinesin-1-driven lipid droplets.
    Cell. 2008 Dec 12;135(6):1098-107 PMID: 19070579
  12. Mass spectrometry-based label-free quantitative proteomics.
    J Biomed Biotechnol. 2010;2010:840518 PMID: 19911078
  13. Targeting the motor regulator Klar to lipid droplets.
    BMC Cell Biol. 2011 Feb 24;12:9 PMID: 21349165
  14. A small predicted stem-loop structure mediates oocyte localization of Drosophila K10 mRNA.
    Development. 1995 Nov;121(11):3809-18 PMID: 8582290
  15. Overexpression of the dynamitin (p50) subunit of the dynactin complex disrupts dynein-dependent maintenance of membrane organelle distribution.
    J Cell Biol. 1997 Oct 20;139(2):469-84 PMID: 9334349
  16. A PAR-1-dependent orientation gradient of dynamic microtubules directs posterior cargo transport in the Drosophila oocyte.
    J Cell Biol. 2011 Jul 11;194(1):121-35 PMID: 21746854
  17. A dynein loading zone for retrograde endosome motility at microtubule plus-ends.
    EMBO J. 2006 Jun 7;25(11):2275-86 PMID: 16688221
  18. Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search.
    Anal Chem. 2002 Oct 15;74(20):5383-92 PMID: 12403597
  19. mRNA localization: gene expression in the spatial dimension.
    Cell. 2009 Feb 20;136(4):719-30 PMID: 19239891
  20. A'-form RNA helices are required for cytoplasmic mRNA transport in Drosophila.
    Nat Struct Mol Biol. 2010 Jun;17(6):703-9 PMID: 20473315
  21. Drosophila Lissencephaly-1 functions with Bic-D and dynein in oocyte determination and nuclear positioning.
    Nat Cell Biol. 1999 Nov;1(7):444-9 PMID: 10559989
  22. A Cdk5-dependent switch regulates Lis1/Ndel1/dynein-driven organelle transport in adult axons.
    J Neurosci. 2011 Nov 23;31(47):17207-19 PMID: 22114287
  23. Mechanisms of dendritic mRNA transport and its role in synaptic tagging.
    EMBO J. 2011 Aug 31;30(17):3540-52 PMID: 21878995
  24. Dynein light intermediate chain: an essential subunit that contributes to spindle checkpoint inactivation.
    Mol Biol Cell. 2008 Nov;19(11):4918-29 PMID: 18799620
  25. Probability-based protein identification by searching sequence databases using mass spectrometry data.
    Electrophoresis. 1999 Dec;20(18):3551-67 PMID: 10612281
  26. Dynactin increases the processivity of the cytoplasmic dynein motor.
    Nat Cell Biol. 2000 Jan;2(1):20-4 PMID: 10620802
  27. NudF, a nuclear migration gene in Aspergillus nidulans, is similar to the human LIS-1 gene required for neuronal migration.
    Mol Biol Cell. 1995 Mar;6(3):297-310 PMID: 7612965
  28. gurken and the I factor retrotransposon RNAs share common localization signals and machinery.
    Dev Cell. 2005 Jul;9(1):51-62 PMID: 15992540
  29. The role of the lissencephaly protein Pac1 during nuclear migration in budding yeast.
    J Cell Biol. 2003 Feb 3;160(3):355-64 PMID: 12566428
  30. An Egalitarian-BicaudalD complex is essential for oocyte specification and axis determination in Drosophila.
    Genes Dev. 1997 Feb 15;11(4):423-35 PMID: 9042857
  31. LIS1 RNA interference blocks neural stem cell division, morphogenesis, and motility at multiple stages.
    J Cell Biol. 2005 Sep 12;170(6):935-45 PMID: 16144905
  32. Molecular characterization of the 50-kD subunit of dynactin reveals function for the complex in chromosome alignment and spindle organization during mitosis.
    J Cell Biol. 1996 Feb;132(4):617-33 PMID: 8647893
  33. Yeast two-hybrid vectors and strains.
    Methods Mol Biol. 2001;177:41-84 PMID: 11530616
  34. High-resolution imaging reveals indirect coordination of opposite motors and a role for LIS1 in high-load axonal transport.
    J Cell Biol. 2011 Oct 17;195(2):193-201 PMID: 22006948
  35. Isolation of a Miller-Dieker lissencephaly gene containing G protein beta-subunit-like repeats.
    Nature. 1993 Aug 19;364(6439):717-21 PMID: 8355785
  36. Mammalian Golgi-associated Bicaudal-D2 functions in the dynein-dynactin pathway by interacting with these complexes.
    EMBO J. 2001 Aug 1;20(15):4041-54 PMID: 11483508
  37. Guidance of bidirectional motor complexes by mRNA cargoes through control of dynein number and activity.
    Curr Biol. 2006 Jul 25;16(14):1447-52 PMID: 16860745
  38. Roles of the Drosophila NudE protein in kinetochore function and centrosome migration.
    J Cell Sci. 2009 Jun 1;122(Pt 11):1747-58 PMID: 19417004
  39. Regulated offloading of cytoplasmic dynein from microtubule plus ends to the cortex.
    Dev Cell. 2011 May 17;20(5):639-51 PMID: 21571221
  40. Lis1 is an initiation factor for dynein-driven organelle transport.
    J Cell Biol. 2012 Jun 25;197(7):971-82 PMID: 22711696
  41. Microtubule binding by dynactin is required for microtubule organization but not cargo transport.
    J Cell Biol. 2007 Feb 26;176(5):641-51 PMID: 17325206
  42. Dictyostelium LIS1 is a centrosomal protein required for microtubule/cell cortex interactions, nucleus/centrosome linkage, and actin dynamics.
    Mol Biol Cell. 2005 Jun;16(6):2759-71 PMID: 15800059
  43. Dynein anchors its mRNA cargo after apical transport in the Drosophila blastoderm embryo.
    Cell. 2005 Jul 15;122(1):97-106 PMID: 16009136
  44. Regulation of dynein localization and centrosome positioning by Lis-1 and asunder during Drosophila spermatogenesis.
    Development. 2012 Aug;139(16):2945-54 PMID: 22764052
  45. Multiple modes of cytoplasmic dynein regulation.
    Nat Cell Biol. 2012 Feb 29;14(3):224-30 PMID: 22373868
  46. Requirement for phosphorylation and localization of the Bicaudal-D protein in Drosophila oocyte differentiation.
    Cell. 1991 Nov 29;67(5):917-26 PMID: 1959135
  47. Targeted mutagenesis of Lis1 disrupts cortical development and LIS1 homodimerization.
    Proc Natl Acad Sci U S A. 2001 May 22;98(11):6429-34 PMID: 11344260
  48. Lis1 acts as a "clutch" between the ATPase and microtubule-binding domains of the dynein motor.
    Cell. 2012 Aug 31;150(5):975-86 PMID: 22939623
  49. Egalitarian is a selective RNA-binding protein linking mRNA localization signals to the dynein motor.
    Genes Dev. 2009 Jul 1;23(13):1546-58 PMID: 19515976
  50. LIS1, CLIP-170's key to the dynein/dynactin pathway.
    Mol Cell Biol. 2002 May;22(9):3089-102 PMID: 11940666
  51. The Drosophila Lissencephaly1 (DLis1) gene is required for nuclear migration.
    Dev Biol. 2000 Oct 1;226(1):57-72 PMID: 10993674
  52. BICD2, dynactin, and LIS1 cooperate in regulating dynein recruitment to cellular structures.
    Mol Biol Cell. 2012 Nov;23(21):4226-41 PMID: 22956769
  53. Ndel1 operates in a common pathway with LIS1 and cytoplasmic dynein to regulate cortical neuronal positioning.
    Neuron. 2004 Oct 14;44(2):263-77 PMID: 15473966
  54. LIS1 and NudE induce a persistent dynein force-producing state.
    Cell. 2010 Apr 16;141(2):304-14 PMID: 20403325
  55. Lis1 and doublecortin function with dynein to mediate coupling of the nucleus to the centrosome in neuronal migration.
    J Cell Biol. 2004 Jun 7;165(5):709-21 PMID: 15173193
  56. Role of dynein, dynactin, and CLIP-170 interactions in LIS1 kinetochore function.
    J Cell Biol. 2002 Mar 18;156(6):959-68 PMID: 11889140
  57. How does a hypha grow? The biophysics of pressurized growth in fungi.
    Nat Rev Microbiol. 2011 Jun 06;9(7):509-18 PMID: 21643041
  58. Evidence for a role of CLIP-170 in the establishment of metaphase chromosome alignment.
    J Cell Biol. 1998 May 18;141(4):849-62 PMID: 9585405
  59. Functional interplay between LIS1, NDE1 and NDEL1 in dynein-dependent organelle positioning.
    J Cell Sci. 2010 Jan 15;123(Pt 2):202-12 PMID: 20048338
  60. Dynamic association of cytoplasmic dynein heavy chain 1a with the Golgi apparatus and intermediate compartment.
    J Cell Sci. 1999 Dec;112 ( Pt 24):4673-85 PMID: 10574715
  61. A LIS1/NUDEL/cytoplasmic dynein heavy chain complex in the developing and adult nervous system.
    Neuron. 2000 Dec;28(3):681-96 PMID: 11163259
  62. Tracking the ends: a dynamic protein network controls the fate of microtubule tips.
    Nat Rev Mol Cell Biol. 2008 Apr;9(4):309-22 PMID: 18322465
  63. A model for random sampling and estimation of relative protein abundance in shotgun proteomics.
    Anal Chem. 2004 Jul 15;76(14):4193-201 PMID: 15253663
  64. Conformational changes in CLIP-170 regulate its binding to microtubules and dynactin localization.
    J Cell Biol. 2004 Sep 27;166(7):1003-14 PMID: 15381688
  65. Polar transport in the Drosophila oocyte requires Dynein and Kinesin I cooperation.
    Curr Biol. 2002 Dec 10;12(23):1971-81 PMID: 12477385
  66. The cytoplasmic dynein and kinesin motors have interdependent roles in patterning the Drosophila oocyte.
    Curr Biol. 2002 Dec 10;12(23):1982-91 PMID: 12477386
  67. Building complexity: an in vitro study of cytoplasmic dynein with in vivo implications.
    Curr Biol. 2005 Dec 6;15(23):2075-85 PMID: 16332532
  68. A statistical model for identifying proteins by tandem mass spectrometry.
    Anal Chem. 2003 Sep 1;75(17):4646-58 PMID: 14632076
  69. Live imaging of Drosophila brain neuroblasts reveals a role for Lis1/dynactin in spindle assembly and mitotic checkpoint control.
    Mol Biol Cell. 2005 Nov;16(11):5127-40 PMID: 16107559
  70. Bicaudal-D and its role in cargo sorting by microtubule-based motors.
    Biochem Soc Trans. 2009 Oct;37(Pt 5):1066-71 PMID: 19754453
  71. Universal sample preparation method for proteome analysis.
    Nat Methods. 2009 May;6(5):359-62 PMID: 19377485
  72. Regulation of cytoplasmic dynein behaviour and microtubule organization by mammalian Lis1.
    Nat Cell Biol. 2000 Nov;2(11):767-75 PMID: 11056530
  73. The microtubule plus-end localization of Aspergillus dynein is important for dynein-early-endosome interaction but not for dynein ATPase activation.
    J Cell Sci. 2010 Oct 15;123(Pt 20):3596-604 PMID: 20876661
  74. Genomic deletions of the Drosophila melanogaster Hsp70 genes.
    Genetics. 2004 Nov;168(3):1467-76 PMID: 15579699
  75. Conserved signals and machinery for RNA transport in Drosophila oogenesis and embryogenesis.
    Nature. 2001 Dec 6;414(6864):611-6 PMID: 11740552
  76. Rapid effects of acute anoxia on spindle kinetochore interactions activate the mitotic spindle checkpoint.
    J Cell Sci. 2007 Aug 15;120(Pt 16):2807-18 PMID: 17652159
  77. Single-molecule assays reveal that RNA localization signals regulate dynein-dynactin copy number on individual transcript cargoes.
    Nat Cell Biol. 2012 Feb 26;14(4):416-23 PMID: 22366687
  78. Regulation of the processivity and intracellular localization of Saccharomyces cerevisiae dynein by dynactin.
    Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5669-74 PMID: 19293377
  79. Egalitarian binds dynein light chain to establish oocyte polarity and maintain oocyte fate.
    Nat Cell Biol. 2004 May;6(5):427-35 PMID: 15077115
  80. A genome-wide RNAi screen to dissect centriole duplication and centrosome maturation in Drosophila.
    PLoS Biol. 2008 Sep 16;6(9):e224 PMID: 18798690
  81. Subcellular mRNA localization in animal cells and why it matters.
    Science. 2009 Nov 27;326(5957):1212-6 PMID: 19965463
  82. Lis1/dynactin regulates metaphase spindle orientation in Drosophila neuroblasts.
    Dev Biol. 2008 Jul 1;319(1):1-9 PMID: 18485341
  83. Functional coordination of three mitotic motors in Drosophila embryos.
    Mol Biol Cell. 2000 Jan;11(1):241-53 PMID: 10637305
  84. Axonal mRNA localization and local protein synthesis in nervous system assembly, maintenance and repair.
    Nat Rev Neurosci. 2012 Apr 13;13(5):308-24 PMID: 22498899
  85. Determinants of S. cerevisiae dynein localization and activation: implications for the mechanism of spindle positioning.
    Curr Biol. 2003 Mar 4;13(5):364-72 PMID: 12620184
  86. Streptavidin aptamers: affinity tags for the study of RNAs and ribonucleoproteins.
    RNA. 2001 Apr;7(4):632-41 PMID: 11345441
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.201211052
Published
2013-08-05
Pages
479-94
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC3734092
Subset
IM
Grants
Medical Research Council · MC_U105178790 · United Kingdom
Medical Research Council · U105178790 · United Kingdom
Wellcome Trust · 081849 · United Kingdom
Wellcome Trust · United Kingdom
Wellcome Trust · 092076 · United Kingdom
External Links
PubMed source DOI: 10.1083/jcb.201211052 PMC Full Text View journal details
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