In situ localization and in vitro induction of plant COPI-coated vesicles

被引:163
作者
Pimpl, P
Movafeghi, A
Coughlan, S
Denecke, J
Hillmer, S
Robinson, DG [1 ]
机构
[1] Univ Gottingen, Albrecht Von Haller Inst Plant Sci, Dept Struct Cell Physiol, Gottingen, Germany
[2] AG Biotechnol, Dupont Expt Stn, Wilmington, DE 19880 USA
[3] Univ Leeds, Leeds Inst Plant Biotechnol & Agr, Leeds LS2 9JT, W Yorkshire, England
关键词
D O I
10.1105/tpc.12.11.2219
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Coat protein (COP)-coated vesicles have been shown to mediate protein transport through early steps of the secretory pathway in yeast and mammalian cells. Here, we attempt to elucidate their role in vesicular trafficking of plant cells, using a combined biochemical and ultrastructural approach. Immunogold labeling of cryosections revealed that COPI proteins are localized to microvesicles surrounding or budding from the Golgi apparatus. COPI-coated buds primarily reside on the cia-face of the Golgi stack. In addition, COPI and Arf1p show predominant labeling of the cis-Golgi stack, gradually diminishing toward the trans-Golgi stack. In vitro COPI-coated vesicle induction experiments demonstrated that Arf1p as well as coatomer could be recruited from cauliflower cytosol onto mixed endoplasmic reticulum (ER)/Golgi membranes. Binding of Arf1p and coatomer is inhibited by brefeldin A, underlining the specificity of the recruitment mechanism. In vitro vesicle budding was confirmed by identification of COPI-coated vesicles through immunogold negative staining in a fraction purified from isopycnic sucrose gradient centrifugation. Similar in vitro induction experiments with tobacco ER/Golgi membranes prepared from transgenic plants overproducing barley alpha -amylase-HDEL yielded a COPI-coated vesicle fraction that contained ol-amylase as well as calreticulin.
引用
收藏
页码:2219 / 2235
页数:17
相关论文
共 75 条
  • [1] The structure and function of the Golgi apparatus: a hundred years of questions
    Andreeva, AV
    Kutuzov, MA
    Evans, DE
    Hawes, CR
    [J]. JOURNAL OF EXPERIMENTAL BOTANY, 1998, 49 (325) : 1281 - 1291
  • [2] SEQUENTIAL COUPLING BETWEEN COPII AND COPI VESICLE COATS IN ENDOPLASMIC-RETICULUM TO GOLGI TRANSPORT
    ARIDOR, M
    BANNYKH, SI
    ROWE, T
    BALCH, WE
    [J]. JOURNAL OF CELL BIOLOGY, 1995, 131 (04) : 875 - 893
  • [3] Cargo selection by the COPII budding machinery during export from the ER
    Aridor, M
    Weissman, J
    Bannykh, S
    Nuoffer, C
    Balch, WE
    [J]. JOURNAL OF CELL BIOLOGY, 1998, 141 (01) : 61 - 70
  • [4] Selective transport of cargo between the endoplasmic reticulum and Golgi compartments
    Bannykh, SI
    Balch, WE
    [J]. HISTOCHEMISTRY AND CELL BIOLOGY, 1998, 109 (5-6) : 463 - 475
  • [5] BARLOWE C, 1993, J BIOL CHEM, V268, P873
  • [6] COPII - A MEMBRANE COAT FORMED BY SEC PROTEINS THAT DRIVE VESICLE BUDDING FROM THE ENDOPLASMIC-RETICULUM
    BARLOWE, C
    ORCI, L
    YEUNG, T
    HOSOBUCHI, M
    HAMAMOTO, S
    SALAMA, N
    REXACH, MF
    RAVAZZOLA, M
    AMHERDT, M
    SCHEKMAN, R
    [J]. CELL, 1994, 77 (06) : 895 - 907
  • [7] SEC12 ENCODES A GUANINE-NUCLEOTIDE-EXCHANGE FACTOR ESSENTIAL FOR TRANSPORT VESICLE BUDDING FROM THE ER
    BARLOWE, C
    SCHEKMAN, R
    [J]. NATURE, 1993, 365 (6444) : 347 - 349
  • [8] Characterization of AtSEC12 and AtSAR1 - Proteins likely involved in endoplasmic reticulum and Golgi transport
    BarPeled, M
    Raikhel, NV
    [J]. PLANT PHYSIOLOGY, 1997, 114 (01) : 315 - 324
  • [9] Stacks on tracks: the plant Golgi apparatus traffics on an actin/ER network
    Boevink, P
    Oparka, K
    Cruz, SS
    Martin, B
    Betteridge, A
    Hawes, C
    [J]. PLANT JOURNAL, 1998, 15 (03) : 441 - 447
  • [10] BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3