Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1992 Aug 1;89(15):7242–7246. doi: 10.1073/pnas.89.15.7242

Brefeldin A affects early events but does not affect late events along the exocytic pathway in pancreatic acinar cells.

L C Hendricks 1, S L McClanahan 1, G E Palade 1, M G Farquhar 1
PMCID: PMC49682  PMID: 1496018

Abstract

Brefeldin A (BFA) blocks protein export from the endoplasmic reticulum (ER) to Golgi complex and causes dismantling of the Golgi complex with relocation of resident Golgi proteins to the ER in some cultured cells. It is not known whether later steps in the secretory process are affected. We previously have shown that in BFA-treated rat pancreatic lobules, there is no detectable relocation of Golgi proteins to the ER and, although Golgi cisternae are rapidly dismantled, clusters of small smooth vesicles consisting of both bona fide Golgi remnants and associated vesicular carriers persist even with prolonged BFA exposure. We now report the effects of BFA on transport of proteins through the secretory pathway in exocrine pancreatic cells; we pulse-labeled pancreatic lobules with [35S]methionine and then chased for various times before adding BFA. When BFA was added at pulse, treated lobules released less than 10% of radioactive protein in comparison with controls, regardless of whether or not the lobule cultures were stimulated with carbamoylcholine. However, when lobules were pulsed and then chased for 30, 45, or 60 min before BFA addition, the amount of labeled protein released was comparable in both BFA-treated and untreated cultures. Furthermore, the kinetics and amounts of basal and carbamoylcholine-stimulated release of unlabeled alpha-amylase from storage in zymogen granules were similar in both control and BFA-treated lobules. Therefore, in the rat pancreas, BFA blocks ER to Golgi transport but does not affect later stages along the secretory pathway, including intra-Golgi transport, exit from the Golgi complex, formation and concentration of secretory granules, and exocytosis.

Full text

PDF
7242

Images in this article

7245Image
on p.7245

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ceska M., Hultman E., Ingelman B. G. A new method for determination of alpha-amylase. Experientia. 1969 May 15;25(5):555–556. doi: 10.1007/BF01900818. [DOI] [PubMed] [Google Scholar]
  2. Doms R. W., Russ G., Yewdell J. W. Brefeldin A redistributes resident and itinerant Golgi proteins to the endoplasmic reticulum. J Cell Biol. 1989 Jul;109(1):61–72. doi: 10.1083/jcb.109.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fujiwara T., Oda K., Yokota S., Takatsuki A., Ikehara Y. Brefeldin A causes disassembly of the Golgi complex and accumulation of secretory proteins in the endoplasmic reticulum. J Biol Chem. 1988 Dec 5;263(34):18545–18552. [PubMed] [Google Scholar]
  4. Hunziker W., Whitney J. A., Mellman I. Selective inhibition of transcytosis by brefeldin A in MDCK cells. Cell. 1991 Nov 1;67(3):617–627. doi: 10.1016/0092-8674(91)90535-7. [DOI] [PubMed] [Google Scholar]
  5. Jamieson J. D., Palade G. E. Intracellular transport of secretory proteins in the pancreatic exocrine cell. II. Transport to condensing vacuoles and zymogen granules. J Cell Biol. 1967 Aug;34(2):597–615. doi: 10.1083/jcb.34.2.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jamieson J. D., Palade G. E. Intracellular transport of secretory proteins in the pancreatic exocrine cell. IV. Metabolic requirements. J Cell Biol. 1968 Dec;39(3):589–603. doi: 10.1083/jcb.39.3.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jamieson J. D., Palade G. E. Synthesis, intracellular transport, and discharge of secretory proteins in stimulated pancreatic exocrine cells. J Cell Biol. 1971 Jul;50(1):135–158. doi: 10.1083/jcb.50.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kato S., Ito S., Noguchi T., Naito H. Effects of brefeldin A on the synthesis and secretion of egg white proteins in primary cultured oviduct cells of laying Japanese quail (Coturnix coturnix japonica). Biochim Biophys Acta. 1989 Apr 25;991(1):36–43. doi: 10.1016/0304-4165(89)90025-1. [DOI] [PubMed] [Google Scholar]
  9. Ktistakis N. T., Roth M. G., Bloom G. S. PtK1 cells contain a nondiffusible, dominant factor that makes the Golgi apparatus resistant to brefeldin A. J Cell Biol. 1991 Jun;113(5):1009–1023. doi: 10.1083/jcb.113.5.1009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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 Mar 10;56(5):801–813. doi: 10.1016/0092-8674(89)90685-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lippincott-Schwartz J., Yuan L., Tipper C., Amherdt M., Orci L., Klausner R. D. Brefeldin A's effects on endosomes, lysosomes, and the TGN suggest a general mechanism for regulating organelle structure and membrane traffic. Cell. 1991 Nov 1;67(3):601–616. doi: 10.1016/0092-8674(91)90534-6. [DOI] [PubMed] [Google Scholar]
  12. Magner J. A., Papagiannes E. Blockade by brefeldin A of intracellular transport of secretory proteins in mouse pituitary cells: effects on the biosynthesis of thyrotropin and free alpha-subunits. Endocrinology. 1988 Mar;122(3):912–920. doi: 10.1210/endo-122-3-912. [DOI] [PubMed] [Google Scholar]
  13. Misumi Y., Misumi Y., Miki K., Takatsuki A., Tamura G., Ikehara Y. Novel blockade by brefeldin A of intracellular transport of secretory proteins in cultured rat hepatocytes. J Biol Chem. 1986 Aug 25;261(24):11398–11403. [PubMed] [Google Scholar]
  14. Oda K., Fujiwara T., Ikehara Y. Brefeldin A arrests the intracellular transport of viral envelope proteins in primary cultured rat hepatocytes and HepG2 cells. Biochem J. 1990 Jan 1;265(1):161–167. doi: 10.1042/bj2650161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Oda K., Nishimura Y. Brefeldin A inhibits the targeting of cathepsin D and cathepsin H to lysosomes in rat hepatocytes. Biochem Biophys Res Commun. 1989 Aug 30;163(1):220–225. doi: 10.1016/0006-291x(89)92124-4. [DOI] [PubMed] [Google Scholar]
  16. Palade G. Intracellular aspects of the process of protein synthesis. Science. 1975 Aug 1;189(4200):347–358. doi: 10.1126/science.1096303. [DOI] [PubMed] [Google Scholar]
  17. Sandvig K., Prydz K., Hansen S. H., van Deurs B. Ricin transport in brefeldin A-treated cells: correlation between Golgi structure and toxic effect. J Cell Biol. 1991 Nov;115(4):971–981. doi: 10.1083/jcb.115.4.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Saraste J., Palade G. E., Farquhar M. G. Temperature-sensitive steps in the transport of secretory proteins through the Golgi complex in exocrine pancreatic cells. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6425–6429. doi: 10.1073/pnas.83.17.6425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Scheele G. A., Palade G. E., Tartakoff A. M. Cell fractionation studies on the guinea pig pancreas. Redistribution of exocrine proteins during tissue homogenization. J Cell Biol. 1978 Jul;78(1):110–130. doi: 10.1083/jcb.78.1.110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Scheele G. Pancreatic lobules in the in vitro study of pancreatic acinar cell function. Methods Enzymol. 1983;98:17–28. doi: 10.1016/0076-6879(83)98135-1. [DOI] [PubMed] [Google Scholar]
  21. Scheele G., Tartakoff A. Exit of nonglycosylated secretory proteins from the rough endoplasmic reticulum is asynchronous in the exocrine pancreas. J Biol Chem. 1985 Jan 25;260(2):926–931. [PubMed] [Google Scholar]
  22. Tartakoff A. M. Temperature and energy dependence of secretory protein transport in the exocrine pancreas. EMBO J. 1986 Jul;5(7):1477–1482. doi: 10.1002/j.1460-2075.1986.tb04385.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tartakoff A., Vassalli P., Détraz M. Comparative studies of intracellular transport of secretory proteins. J Cell Biol. 1978 Dec;79(3):694–707. doi: 10.1083/jcb.79.3.694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Thomas B. J., Rothstein R. Sex, maps, and imprinting. Cell. 1991 Jan 11;64(1):1–3. doi: 10.1016/0092-8674(91)90199-9. [DOI] [PubMed] [Google Scholar]
  25. Ulmer J. B., Palade G. E. Effects of Brefeldin A on the Golgi complex, endoplasmic reticulum and viral envelope glycoproteins in murine erythroleukemia cells. Eur J Cell Biol. 1991 Feb;54(1):38–54. [PubMed] [Google Scholar]
  26. Ulmer J. B., Palade G. E. Targeting and processing of glycophorins in murine erythroleukemia cells: use of brefeldin A as a perturbant of intracellular traffic. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6992–6996. doi: 10.1073/pnas.86.18.6992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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 Nov 1;67(3):591–600. doi: 10.1016/0092-8674(91)90533-5. [DOI] [PubMed] [Google Scholar]
  28. Zizi M., Fisher R. S., Grillo F. G. Formation of cation channels in planar lipid bilayers by brefeldin A. J Biol Chem. 1991 Oct 5;266(28):18443–18445. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES