Download PDF
Plant Biol (Stuttg) 2007; 9(4): 489-501
DOI: 10.1055/s-2006-924759
Research Paper

Georg Thieme Verlag Stuttgart KG · New York

Brefeldin A Action and Recovery in Chlamydomonas are Rapid and Involve Fusion and Fission of Golgi Cisternae

E. Hummel[*] 1 , R. Schmickl[*] 1 , G. Hinz1 , S. Hillmer1 , D. G. Robinson1
  • 1Department of Cell Biology, Heidelberg Institute for Plant Sciences (HIP), University of Heidelberg, Im Neuenheimer Feld 230, 69120 Heidelberg, Germany
Further Information

Publication History

Received: July 4, 2006

Accepted: October 18, 2006

Publication Date:
15 February 2007 (online)

   Permissions and Reprints

Abstract

Chlamydomonas noctigama has a non-motile Golgi apparatus consisting of several Golgi stacks adjacent to transitional ER. These domains are characterized by vesicle-budding profiles and the lack of ribosomes on the side of the ER proximal to the Golgi stacks. Immunogold labelling confirms the presence of COPI-proteins at the periphery of the Golgi stacks, and COPII-proteins at the ER-Golgi interface. After addition of BFA (10 µg/ml) a marked increase in the number of vesicular profiles lying between the ER and the Golgi stacks is seen. Serial sections of cells do not provide any evidence for the existence of tubular connections between the ER and the Golgi stacks, supporting the notion that COPI- but not COPII-vesicle production is affected by BFA. The fusion of COPII-vesicles at the cis-Golgi apparatus apparently requires the presence of retrograde COPI-vesicles. After 15 min the cisternae of neighbouring Golgi stacks begin to fuse forming “mega-Golgis”, which gradually curl before fragmenting into clusters of vesicles and tubules. These are surrounded by the transitional ER on which vesicle-budding profiles are still occasionally visible. Golgi remnants continue to survive for several hours and do not completely disappear. Washing out BFA leads to a very rapid reassembly of Golgi cisternae. At first, clusters of vesicles are seen adjacent to transitional ER, then “mini Golgis” are seen whose cisternae grow in length and number to produce “mega Golgis”. These structures then divide by vertical fission to produce Golgi stacks of normal size and morphology roughly 60 min after drug wash-out.

Key words

Brefeldin A - Chlamydomonas - COP-vesicles - Golgi fission - Golgi fusion.

References

  • 1 Altan-Bonnet N., Sougrat R., Lippincott-Schwartz J.. Molecular basis for Golgi maintenance and biogenesis.  Current Opinions in Cell Biology. (2004);  16 364-372
    PubMedGoogle Scholar
  • 68 Amrhein N., Filner P.. Adenosine 3′:5′-cyclic monophosphate in Chlamydomonas reinhardtii: isolation and characterization.  Proceedings of the National Academy of Sciences of the USA. (1973);  70 1099-1103
    CrossrefPubMedGoogle Scholar
  • 2 Aniento F., Matsuoka K., Robinson D. G.. ER to Golgi transport: the COPII pathway. Robinson, D. G., ed. The Plant Endoplasmic Reticulum, Plant Cell Monographs, Vol. 4. Heidelberg; Springer Verlag (2006): 99-124
    Google Scholar
  • 3 Balch W. E.. Vesicle traffic in vitro.  Cell. (2004);  116 17-19
    CrossrefPubMedGoogle Scholar
  • 4 Bannykh S. I., Rowe T., Balch W. E.. The organization of endoplasmic reticulum export complexes.  Journal of Cell Biology. (1996);  135 19-35
    CrossrefPubMedGoogle Scholar
  • 5 Becker B., Hickisch A.. Inhibition of contractile vacuole function by brefeldin A.  Plant Cell Physiology. (2005);  46 201-212
    CrossrefPubMedGoogle Scholar
  • 6 Becker B., Melkonian M.. The secretory pathway of protists: spatial and functional organization and evolution.  Microbiology Reviews. (1996);  60 697-721
    PubMedGoogle Scholar
  • 7 Benchimol M., Ribiero K. C., Mariante R. M., Alderete J. F.. Structure and division of the Golgi complex in Trichomonas vaginalis and Tritrichomonas foetus.  European Journal of Cell Biology. (2001);  80 593-607
    CrossrefPubMedGoogle Scholar
  • 8 Bevis B. J., Hammond A. T., Reinke C. A., Glick B. S.. De novo formation of transitional ER sites and Golgi structures in Pichia pastoris.  Nature Cell Biology. (2002);  4 750-756
    CrossrefPubMedGoogle Scholar
  • 9 Boevink P., Oparka K., Santa Cruz S., Martin B., Betteridge A., Hawes C.. Stacks on tracks: the plant Golgi apparatus traffics on an actin/ER network.  The Plant Journal. (1998);  15 441-447
    CrossrefPubMedGoogle Scholar
  • 69 Bonfanti L., Mironov A., Martinez-Menarguez J., Martella O., Fusella A., Baldassare M., Buccione R., Geuze H., Mironov A., Luini A.. Procollagen traverses the Golgi stack without leaving the lumen of cisternae: evidence for cisternal maturation.  Cell. (1998);  95 993-1003
    CrossrefPubMedGoogle Scholar
  • 10 Bonifacino J. S., Glick B. S.. The mechanisms of vesicle budding and fusion.  Cell. (2004);  116 153-166
    CrossrefPubMedGoogle Scholar
  • 11 Brandizzi F., Snap E., Roberts A., Lippincott-Schwartz J., Hawes C.. Membrane protein transport between the ER and Golgi in tobacco leaves is energy dependent but cytoskeleton independent: evidence from selective photobleaching.  Plant Cell. (2002);  14 1293-1309
    CrossrefPubMedGoogle Scholar
  • 12 Cox R., Mason-Gamer R. J., Jackson C. L., Segev N.. Phylogenetic analysis of Sec7-domain-containing Arf nucleotide exchangers.  Molecular Biology of the Cell. (2004);  15 1487-1505
    CrossrefPubMedGoogle Scholar
  • 13 daSilva L. L., Snapp E. L., Denecke J., Lippincott-Schwartz J., Hawes C., Brandizzi F.. Endoplasmic reticulum export sites and Golgi bodies behave as single mobile secretory units in plant cells.  Plant Cell. (2004);  16 1753-1771
    CrossrefPubMedGoogle Scholar
  • 14 Dairman M., Donofrio N., Domozych D. S.. The effects of brefeldin A upon the Golgi apparatus of the green algal flagellate, Gloeomonas kupfferi.  Journal of Experimental Botany. (1995);  46 181-186
    CrossrefPubMedGoogle Scholar
  • 15 Donaldson J. G., Honda A., Weigert R.. Multiple activities for Arf1 at the Golgi complex.  Biochimica et Biophysica Acta. (2005);  1744 364-373
    PubMedGoogle Scholar
  • 16 Domozych D. S.. Disruption of the Golgi apparatus and secretory mechanism in the desmid, Closterium acerosum by brefeldin A.  Journal of Experimental Botany. (1999);  50 1323-1330
    CrossrefPubMedGoogle Scholar
  • 17 Falk H., Kleinig H.. Feinbau und Carotinoide von Tribonema (Xantophyceae).  Archiv für Mikrobiologie. (1968);  61 347-362
    PubMedGoogle Scholar
  • 18 Geldner N., Anders N., Wolters H., Keicher J., Kornberger W., Muller P., Delbarre A., Ueda T., Nakano A., Jurgens G.. The Arabidopsis GNOM ARF‐GEF mediates endosomal recycling, auxin transport, and auxin-dependent plant growth.  Cell. (2003);  112 219-230
    CrossrefPubMedGoogle Scholar
  • 19 Geldner N., Richter S., Vieten A., Marquardt S., Torres-Ruiz R.A., Mayer U., Jürgens G.. Partial loss-of-function alleles reveal a role for GNOM in auxin transport-related, post-embryonic development of Arabidopsis.  Development. (2004);  131 389-400
    PubMedGoogle Scholar
  • 20 Geldner N.. The plant endosomal system - its structure and role in signal transduction and plant development.  Planta. (2004);  219 547-560
    PubMedGoogle Scholar
  • 70 Glick B. S., Malhotra V. S.. The curious status of the Golgi apparatus.  Cell. (1998);  95 883-889
    CrossrefPubMedGoogle Scholar
  • 21 Gruber H. E., Rosario B.. Ultrastructure of the Golgi apparatus and contractile vacuole in Chlamydomonas reinhardtii. .  Cytologica. (1979);  44 505-526
    PubMedGoogle Scholar
  • 22 Haller K., Fabry S.. Brefeldin A affects synthesis and integrity of a eukaryotic flagellum.  Biochemistry and Biophysical Research Communications. (1998);  242 597-601
    PubMedGoogle Scholar
  • 23 Hawes C.. The plant Golgi apparatus.  New Phytologist. (2005);  165 29-44
    CrossrefPubMedGoogle Scholar
  • 24 Hawes C., Satiat-Jeunemaitre B.. The plant Golgi apparatus - going with the flow.  Biochimica et Biophysica Acta. (2005);  1744 93-107
    PubMedGoogle Scholar
  • 25 He C. Y., Ho H. H., Malsam J., Chalouni C., West C. M., Ullu E., Toomre D., Warren G.. Golgi duplication in Trypanosoma brucei.  Journal of Cell Biology. (2004);  165 313-321
    CrossrefPubMedGoogle Scholar
  • 26 Jackson C. L., Casanova J. E.. Turning on Arf: the sec7 familiy of guanine nucleotide exchange factors.  Trends in Cell Biology. (2000);  121 317-333
    PubMedGoogle Scholar
  • 27 Jürgens G.. Membrane trafficking in plants.  Annual Reviews in Cell Biology. (2004);  20 481-504
    PubMedGoogle Scholar
  • 28 Klausner R. D., Donaldson J. G., Lippincott-Schwartz J.. Brefeldin A: insights into the control of membrane traffic and organelle structure.  Journal of Cell Biology. (1992);  116 1071-1080
    CrossrefPubMedGoogle Scholar
  • 72 Ladinsky M. S., Mastronarde D. N., McIntosh J. R., Howell K. E., Staehelin L. A.. Golgi structure in three dimensions: functional insights from the normal rat kidney cell.  Journal of Cell Biology. (1999);  114 1135-1149
    PubMedGoogle Scholar
  • 73 Lamandé S. R., Bateman J. F.. Procollagen folding and assembly: the role of endoplasmic reticulum enzymes and molecular chaperones.  Seminars in Cell and Developmental Biology. (1999);  10 455-464
    CrossrefPubMedGoogle Scholar
  • 29 Lee M. C. S., Miller E. A., Goldberg J., Orci L., Schekman R.. Bi-directional protein transport between the ER and Golgi.  Annual Reviews in Cell and Developmental Biology. (2004);  20 87-123
    PubMedGoogle Scholar
  • 30 Lippincott-Schwartz J., Yuan L. C., Bonifacino J. S., Klausner R. D.. Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: evidence for membrane cycling from Golgi to ER.  Cell. (1989);  56 801-813
    CrossrefPubMedGoogle Scholar
  • 31 Lucic V.. Structural studies by electron tomography: from cells to molecules.  Annual Reviews in Biochemistry. (2005);  74 833-865
    CrossrefPubMedGoogle Scholar
  • 74 Meindl U., Lancelle S., Hepler P. K.. Vesicle production and fusion during lobe formation in Micrasterias visualized by high-pressure freeze fixation.  Protoplasma. (1992);  170 104-114
    CrossrefPubMedGoogle Scholar
  • 32 Mironov A. A., Beznoussenko G. V., Trucco A., Lupetti P., Smith J. D., Geerts W. J., Koster A. J., Burger K. N., Martone M. E., Deerinck T. J., Ellismann M. H., Luini A.. ER‐to-Golgi carriers arise through en bloc protusion and multistage maturation of specialized ER exit domains.  Developmental Cell. (2003);  5 583-594
    CrossrefPubMedGoogle Scholar
  • 33 Morré J., Mollenhauer H. H., Bracker C. E.. Origin and continuity of Golgi apparatus. Reinert, J. and Ursprung, H., eds. Origin and Continuity of Cell Organelles. Berlin, Heidelberg, New York; Springer Verlag (1971): 82-126
    Google Scholar
  • 34 Movafeghi. A., Happel N., Pimpl P., Tai G. H., Robinson D. G.. Arabidopsis Sec21 and Sec23 homologs. Probable coat proteins of plant COP-coated vesicles.  Plant Physiology. (1999);  119 1437-1446
    CrossrefPubMedGoogle Scholar
  • 75 Nebenführ A., Gallagher L. A., Dunahay T. G., Frohlick J. A., Mazurkiewicz A. M., Meehl J. B., Staehelin L. A.. Stop-and-go movements of plant Giolgi stacks are mediated by the acto-myosin system.  Plant Physiology. (1999);  121 1127-1141
    CrossrefPubMedGoogle Scholar
  • 35 Nebenführ A., Ritzenthaler C., Robinson D. G.. Brefeldin A: deciphering an enigmatic inhibitor of secretion.  Plant Physiology. (2002);  130 1102-1108
    CrossrefPubMedGoogle Scholar
  • 36 Noguchi T., Watanabe H., Suzuki R.. Effects of brefeldin A on the Golgi apparatus, the nuclear envelope and the endoplasmic reticulum in a green alga, Scenedesmus acutus.  Protoplasma. (1998);  201 202-212
    CrossrefPubMedGoogle Scholar
  • 37 Noguchi T., Watanabe H.. Brefeldin A effects on the trans-Golgi network and Golgi bodies in Botryococcus braunii are not uniform during the cell cycle.  Protoplasma. (1999);  209 193-206
    CrossrefPubMedGoogle Scholar
  • 38 Pelletier L., Stern C. A., Pypaert M., Sheff D., Ngo H. M., Roper N., He C. Y., Hu K., Toomre D., Coppens I., Roos D. S., Joiner K. A., Warren G.. Golgi biogenesis in Toxoplasma gondii.  Nature. (2002);  418 548-552
    PubMedGoogle Scholar
  • 39 Phillipson B., Pimpl P., Pinto da Silva L., Crofts A. J., Taylor P., Movafeghi A., Robinson D. G., Denecke J.. Secretory bulk flow of soluble proteins is efficient and COPII dependent.  Plant Cell. (2001);  13 2005-2020
    CrossrefPubMedGoogle Scholar
  • 40 Pimpl P., Movafeghi A., Coughlan S., Denecke J., Hillmer S., Robinson D. G.. In situ localization and in vitro induction of plant COPI-coated vesicles.  Plant Cell. (2000);  12 2219-2236
    CrossrefPubMedGoogle Scholar
  • 41 Polischuck R. S., Mironov A. A.. Structural aspects of Golgi function.  Cell and Molecular Life Sciences. (2004);  61 146-158
    PubMedGoogle Scholar
  • 43 Presley J. F., Smith C., Hirschberg K., Miller C., Cole N. B., Zaal K. J., Lippincott-Schwartz J.. Golgi membrane dynamics.  Molecular Biology of the Cell. (1998);  7 1617-1626
    PubMedGoogle Scholar
  • 44 Pröschold T., Birger M., Schlösser U. W., Melkonian M.. Molecular phylogeny and taxonomic revision of Chlamydomonas (Chlorophyta). I. Emendation of Chlamydomonas Ehrenberg and Chloromonas Gobi and description of Oogochlamys gen. nov. and Lobochlamys gen. nov.  Protist. (2001);  152 265-300
    CrossrefPubMedGoogle Scholar
  • 45 Renault L., Guibert B., Cherfils J.. Structural snapshots of the mechanism and inhibition of a guanine nucleotide exchange factor.  Nature. (2003);  426 525-530
    PubMedGoogle Scholar
  • 76 Ritzenthaler C., Nebenführ A., Movafeghi A., Stussi-Garaud C., Behnia L., Pimpl P., Staehelin L. A., Robinson D. G.. Reevaluation of the effects of brefeldin A on plant cells using tobacco Bright Yellow 2 cells expressing Golgi-targeted green fluorescent protein and COPI antisera.  Plant Cell. (2002);  14 237-261
    CrossrefPubMedGoogle Scholar
  • 77 Robinson D. G., Kristen U.. Membrane flow via the Golgi apparatus of higher plant cells.  International Review of Cytology. (1982);  77 89-127
    PubMedGoogle Scholar
  • 46 Runions J., Brach T., Kühner S., Hawes C.. Photoactivation of GFP reveals protein dynamics within the endoplasmic reticulum membrane.  Journal of Experimental Botany. (2006);  57 43-50
    PubMedGoogle Scholar
  • 47 Salomon S., Meindl U.. Brefeldin A induces reversible dissociation of the Golgi apparatus in the green alga Micrasterias.  Protoplasma. (1996);  194 231-242
    CrossrefPubMedGoogle Scholar
  • 48 Samaj J., Read N. D., Volkmann D., Menzel D., Baluska F.. The endocytic network in plants.  Trends in Cell Biology. (2005);  15 425-433
    CrossrefPubMedGoogle Scholar
  • 49 Sasso A., de Faria F. P., Iwamura E. S., Correa H.. A three dimensional reconstruction study of the rough ER-Golgi interface in serial thin sections of the pancreatic acinar cell of the rat.  Journal of Cell Science. (1994);  107 517-528
    PubMedGoogle Scholar
  • 78 Satiat-Jeunemaitre B., Hawes C. R.. Redistribution of a Golgi glycoprotein in plant cells treated with brefeldin A.  Journal of Cell Science. (1992);  103 1153-1166
    PubMedGoogle Scholar
  • 50 Satiat-Jeunemaitre B., Cole L., Bourett T., Howard R., Hawes C.. Brefeldin A effects in plant and fungal cells: something new about vesicle trafficking?.  Journal of Microscopy. (1996);  181 162-177
    PubMedGoogle Scholar
  • 51 Scheel J., Pepperkok R., Lowe M., Griffiths G., Kreis T.. Dissociation of coatomer from membranes is required brefeldin A-induced transfer of Golgi enzymes to the endoplasmic reticulum.  Journal of Cell Biology. (1997);  137 319-333
    CrossrefPubMedGoogle Scholar
  • 52 Schekman R., Orci L.. Coat proteins and vesicle budding.  Science. (1996);  271 1526-1533
    CrossrefPubMedGoogle Scholar
  • 53 Sciaky N., Presley J., Smith C., Zaal K. J., Cole N., Moreira J. E., Terasaki M., Siggia E., Lippincott-Schwartz J.. Golgi tubule traffic and the effects of brefeldin A visualized in living cells.  Journal of Cell Biology. (1997);  139 1137-1155
    CrossrefPubMedGoogle Scholar
  • 54 Staehelin L. A.. The plant ER: a dynamic organelle composed of a large number of discrete functional domains.  The Plant Journal. (1997);  11 1151-1165
    CrossrefPubMedGoogle Scholar
  • 55 Staehelin L. A., Driouich A.. Brefeldin A effects in plants: are different Golgi responses caused by different sites of action?.  Plant Physiology. (1997);  114 401-403
    PubMedGoogle Scholar
  • 56 Stefano G., Renna L., Chatre L., Hanton S. L., Moreau P., Hawes C., Brandizzi F.. In tobacco leaf epidermal cells, the integrity of protein export from the endoplasmic reticulum and of ER export sites depends on active COPI machinery.  The Plant Journal. (2006);  46 95-110
    CrossrefPubMedGoogle Scholar
  • 57 Steinmann T., Geldner N., Grebe M., Mangold S., Jackson C. L., Paris S., Gälweiler L., Palme K., Jürgens G.. Coordinated polar localization of auxin efflux carrier PIN1 by GNOM ARF GEF.  Science. (1999);  286 316-318
    CrossrefPubMedGoogle Scholar
  • 58 Stephens D. J., Pepperkok R.. Imaging of procollagen transports reaveals COPI dependent cargo sorting during ER‐to-Golgi transport in mammalian cells.  Journal of Cell Science. (2002);  115 1149-1160
    PubMedGoogle Scholar
  • 59 Takeuchi M., Ueda T., Sato K., Abe H., Nagata T., Nakano A.. A dominant negative mutant of sar1 GTPase inhibits protein transport from the endoplasmic reticulum to the Golgi apparatus in tobacco and Arabidopsis cultured cells.  The Plant Journal. (2000);  23 517-525
    CrossrefPubMedGoogle Scholar
  • 60 Tanaka Y., Noguchi T.. Cytoskeleton mediating transport between the ER system and the Golgi apparatus in the green alga Scenedesmus acutus.  European Journal of Cell Biology. (2000);  79 750-758
    CrossrefPubMedGoogle Scholar
  • 79 Ward T. H., Polishchuk S., Caplan S., Hirschberg K., Lippincott-Schwarz J.. Maintenance of Golgi structure and function depends on the integrity of ER export.  Journal of Cell Biology. (2001);  155 557-570
    CrossrefPubMedGoogle Scholar
  • 62 Warren G.. Membrane partitioning during cell division.  Annual Reviews of Biochemistry. (1993);  62 323-348
    PubMedGoogle Scholar
  • 63 Wood S. A., Park J. E., Brown W. J.. Brefeldin A causes a microtubule-mediated fusion of the trans Golgi network and early endosomes.  Cell. (1991);  67 591-600
    CrossrefPubMedGoogle Scholar
  • 64 Yang Y. D., Elamawi R., Bubeck J., Pepperkok R., Ritzenthaler C., Robinson D. G.. Dynamics of COPII vesicles and the Golgi apparatus in cultured Nicotiana tabacum BY‐2 cells provides evidence for transient association of Golgi stacks with endoplasmic reticulum exit sites.  Plant Cell. (2005);  17 1513-1531
    CrossrefPubMedGoogle Scholar
  • 65 Zaal K. J. M., Smith C. L., Polishchuk R. S., Altan N., Cole N. B., Ellenberg J., Hirschberg K., Presley J. F., Roberts T. H., Siggia E., Phair R. D., Lippincott-Schwartz J.. Golgi membranes are adsorbed into and reemerge from the ER during mitosis.  Cell. (1999);  99 589-601
    CrossrefPubMedGoogle Scholar
  • 66 Zeuschner D., Geerts W. J. C., van Donselaar E., Humbel B. M., Slot J. W., Koster A., Klumperman J.. Immuno-electron tomography of ER exit sites reveals the existence of free COPII-coated transport carriers.  Nature Cell Biology. (2006);  8 377-383
    CrossrefPubMedGoogle Scholar
  • 67 Zhang Y.-H., Robinson D. G.. The endomembranes of Chlamydomonas reinhardii: a comparison of the wildtype with wall mutants CW2 and CW15.  Protoplasma. (1986);  133 186-194
    CrossrefPubMedGoogle Scholar

1 These authors contributed equally to this paper

D. G. Robinson

Department of Cell Biology
Heidelberg Institute for Plant Sciences (HIP)
University of Heidelberg

Im Neuenheimer Feld 230

69120 Heidelberg

Germany

Email: david.robinson@urz.uni-heidelberg.de

Editor: S. M. Wick

      >