Localization and recycling of gp27 (hp24γ3):: Complex formation with other p24 family members

被引:119
作者
Füllekrug, J
Suganuma, T
Tang, BL
Hong, WJ
Storrie, B
Nilsson, T [1 ]
机构
[1] European Mol Biol Lab, Cell Biol & Cell Biophys Program, D-69117 Heidelberg, Germany
[2] Miyazaki Med Coll, Dept Anat, Miyazaki 8891692, Japan
[3] Inst Mol & Cell Biol, Membrane Biol Lab, Singapore 117609, Singapore
[4] Virginia Polytech Inst & State Univ, Dept Biochem, Blacksburg, VA 24061 USA
关键词
D O I
10.1091/mbc.10.6.1939
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We report here the characterization of gp27 (hp24 gamma(3)), a glycoprotein of the p24 family of small and abundant transmembrane proteins of the secretory pathway. Immunoelectron and confocal scanning microscopy show that at steady state, gp27 localizes to the cis side of the Golgi apparatus. In addition, some gp27 was detected in COPI- and COPII-coated structures throughout the cytoplasm. This indicated cycling that was confirmed in three ways. First, 15 degrees C temperature treatment resulted in accumulation of similar to p27 in pre-Golgi structures colocalizing with anterograde cargo. Second, treatment with brefeldin A caused gp27 to relocate into peripheral structures positive for both KDEL receptor and COPII. Third, microinjection of a dominant negative mutant of Sar1p trapped gp27 in the endoplasmic reticulum (ER) by blocking ER export. Together, this shows that gp27 cycles extensively in the early secretory pathway. Immunoprecipitation and coexpression studies further revealed that a significant fraction of gp27 existed in a hetero-oligomeric complex. Three members of the p24 family, GMP25 (hp24 alpha(2)), p24 (hp24 beta(1)), and p23 (hp24 delta(1)), coprecipitated in what appeared to be stochiometric amounts. This heterocomplex was specific. Immunoprecipitation of p26 (hp24 gamma(4)) failed to coprecipitate GMP25, p24, or p23. Also, very little p26 was found coprecipitating with gp27. A functional requirement for complex formation was suggested at the level of ER export. Transiently expressed gp27 failed to leave the ER unless other p24 family proteins were coexpressed. Comparison of attached oligosaccharides showed that gp27 and GMP25 recycled differentially. Only a very minor portion of GMP25 displayed complex oligosaccharides. In contrast, all of gp27 showed modifications by medial and trans enzymes at steady state. We conclude from these data that a portion of gp27 exists as hetero-oligomeric complexes with GMP25, p24, and p23 and that these complexes are in dynamic equilibrium with individual p24 proteins to allow for differential recycling and distributions.
引用
收藏
页码:1939 / 1955
页数:17
相关论文
共 37 条
  • [1] 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
  • [2] Erv25p, a component of COPII-coated vesicles, forms a complex with Emp24p that is required for efficient endoplasmic reticulum to Golgi transport
    Belden, WJ
    Barlowe, C
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (43) : 26939 - 26946
  • [3] Tmp21 and p24A, two type I proteins enriched in pancreatic microsomal membranes, are members of a protein family involved in vesicular trafficking
    Blum, R
    Feick, P
    Puype, M
    Vandekerckhove, J
    Klengel, R
    Nastainczyk, W
    Schulz, I
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (29) : 17183 - 17189
  • [4] gp25L/emp24/p24 protein family members of the cis-Golgi network bind both COP I and II coatomer
    Dominguez, M
    Dejgaard, K
    Füllekrug, J
    Dahan, S
    Fazel, A
    Paccaud, JP
    Thomas, DY
    Bergeron, JJM
    Nilsson, T
    [J]. JOURNAL OF CELL BIOLOGY, 1998, 140 (04) : 751 - 765
  • [5] ATTACHMENT OF TERMINAL N-ACETYLGLUCOSAMINE TO ASPARAGINE-LINKED OLIGOSACCHARIDES OCCURS IN CENTRAL CISTERNAE OF THE GOLGI STACK
    DUNPHY, WG
    BRANDS, R
    ROTHMAN, JE
    [J]. CELL, 1985, 40 (02) : 463 - 472
  • [6] CAVEOLAE AND SORTING IN THE TRANS-GOLGI NETWORK OF EPITHELIAL-CELLS
    DUPREE, P
    PARTON, RG
    RAPOSO, G
    KURZCHALIA, TV
    SIMONS, K
    [J]. EMBO JOURNAL, 1993, 12 (04) : 1597 - 1605
  • [7] Genes that control the fidelity of endoplasmic reticulum to Golgi transport identified as suppressors of vesicle budding mutations
    ElrodErickson, MJ
    Kaiser, CA
    [J]. MOLECULAR BIOLOGY OF THE CELL, 1996, 7 (07) : 1043 - 1058
  • [8] ISOLATION OF MONOCLONAL-ANTIBODIES SPECIFIC FOR HUMAN C-MYC PROTO-ONCOGENE PRODUCT
    EVAN, GI
    LEWIS, GK
    RAMSAY, G
    BISHOP, JM
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 1985, 5 (12) : 3610 - 3616
  • [9] Characterization of brefeldin A induced vesicular structures containing cycling proteins of the intermediate compartment/cis-Golgi network
    Fullekrug, J
    Sonnichsen, B
    Schafer, U
    Van, PN
    Soling, HD
    Mieskes, G
    [J]. FEBS LETTERS, 1997, 404 (01) : 75 - 81
  • [10] Cloning of a putative ligand for the T1/ST2 receptor
    Gayle, MA
    Slack, JL
    Bonnert, TP
    Renshaw, BR
    Sonoda, G
    Taguchi, T
    Testa, JR
    Dower, SK
    Sims, JE
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (10) : 5784 - 5789