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PMID: 9399951 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Overexpression of Rab3D enhances regulated amylase secretion from pancreatic acini of transgenic mice.

The Journal of clinical investigation ·Vol. 100 ·No. 12 ·1997-12-15 ·Pages 3044-52

Ohnishi H, Samuelson LC, Yule DI, Ernst SA, Williams JA

Abstract

Rab3D, a member of the ras-related GTP-binding protein Rab family, is localized to secretory granules of various exocrine tissues such as acinar cells of the pancreas, chief cells of the stomach, and parotid and lacrimal secretory cells. To elucidate the function of Rab3D in exocytosis, we have generated transgenic mice that over-express Rab3D specifically in pancreatic acinar cells. Hemagglutinin-tagged Rab3D was localized to zymogen granules by immunohistochemistry, and was shown to be present on zymogen granule membranes by Western blotting; both results are similar to previous studies of endogenous Rab3D. Secretion measurements in isolated acinar preparations showed that overexpression of Rab3D enhanced amylase release. Amylase secretion from intact acini of transgenic mice 5 min after 10 pM cholecystokinin octapeptide (CCK) stimulation was enhanced by 160% of control. In streptolysin-O-permeabilized acini of transgenic mice, amylase secretion induced by 100 microM GTP-gamma-S was enhanced by 150%, and 10 microM Ca2+-stimulated amylase secretion was augmented by 206% of that of the control. To further elucidate Rab3D involvement in stimulus-secretion coupling, we examined the effect of CCK on the rate of GTP binding to Rab3D. Stimulation of permeabilized acini with 10 pM CCK increased the incorporation of radiolabeled GTP into HA-tagged Rab3D. These results indicate that overexpression of Rab3D enhances secretagogue-stimulated amylase secretion through both calcium and GTP pathways. We conclude that Rab3D protein on zymogen granules plays a stimulatory role in regulated amylase secretion from pancreatic acini.

MeSH Terms
Amylases/metabolism Animals Bacterial Proteins Calcium/pharmacology Cell Membrane Permeability Cholecystokinin/pharmacology Exocytosis GTP-Binding Proteins/biosynthesis,genetics,metabolism Gene Expression Guanosine 5'-O-(3-Thiotriphosphate)/pharmacology Guanosine Triphosphate/metabolism Hemagglutinins/genetics,metabolism Mice Mice, Inbred C57BL Mice, Transgenic Pancreas/drug effects,enzymology,metabolism Streptolysins/pharmacology rab3 GTP-Binding Proteins
Chemicals
Bacterial Proteins Hemagglutinins Streptolysins streptolysin O Guanosine 5'-O-(3-Thiotriphosphate) Guanosine Triphosphate Cholecystokinin Amylases GTP-Binding Proteins rab3 GTP-Binding Proteins Calcium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Ohnishi H
Department of Physiology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Samuelson L C
Yule D I
Ernst S A
Williams J A
References (43)
43 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Rab proteins in regulated exocytosis.
    Trends Biochem Sci. 1994 Apr;19(4):164-8 PMID: 8016866
  3. Specific expression of an elastase-human growth hormone fusion gene in pancreatic acinar cells of transgenic mice.
    Nature. 1985 Feb 14-20;313(6003):600-2 PMID: 3844051
  4. A simple salting out procedure for extracting DNA from human nucleated cells.
    Nucleic Acids Res. 1988 Feb 11;16(3):1215 PMID: 3344216
  5. GE: a GTP-binding protein mediating exocytosis.
    Annu Rev Physiol. 1990;52:591-606 PMID: 2109965
  6. Amylase release from streptolysin O-permeabilized pancreatic acini.
    Am J Physiol. 1990 Aug;259(2 Pt 1):G157-64 PMID: 1696432
  7. Ca2(+)-dependent amylase secretion from pancreatic acinar cells occurs without activation of phospholipase C linked G-proteins.
    Biochem Biophys Res Commun. 1991 Jan 31;174(2):536-41 PMID: 1899567
  8. Small GTP-binding proteins in vesicular transport.
    Trends Biochem Sci. 1990 Dec;15(12):473-7 PMID: 2077687
  9. A synthetic peptide of the rab3a effector domain stimulates amylase release from permeabilized pancreatic acini.
    Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1656-60 PMID: 1371881
  10. Hydrolysis of GTP by Sec4 protein plays an important role in vesicular transport and is stimulated by a GTPase-activating protein in Saccharomyces cerevisiae.
    Mol Cell Biol. 1992 May;12(5):2017-28 PMID: 1569938
  11. Cloning of a Rab3 isotype predominantly expressed in adipocytes.
    Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5049-52 PMID: 1594612
  12. Epidermal growth factor regulates the exchange rate of guanine nucleotides on p21ras in fibroblasts.
    Mol Cell Biol. 1993 Mar;13(3):1903-10 PMID: 8441421
  13. Rabphilin-3A, a putative target protein for smg p25A/rab3A p25 small GTP-binding protein related to synaptotagmin.
    Mol Cell Biol. 1993 Apr;13(4):2061-8 PMID: 8384302
  14. Mutants of Rab3A analogous to oncogenic Ras mutants. Sensitivity to Rab3A-GTPase activating protein and Rab3A-guanine nucleotide releasing factor.
    J Biol Chem. 1993 May 5;268(13):9410-5 PMID: 8387493
  15. Inhibition of Rab3B expression attenuates Ca(2+)-dependent exocytosis in rat anterior pituitary cells.
    Nature. 1993 Aug 5;364(6437):540-4 PMID: 8393147
  16. Expression and localization of two low molecular weight GTP-binding proteins, Rab8 and Rab10, by epitope tag.
    Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6508-12 PMID: 7688123
  17. rab3-peptide stimulates exocytosis from mast cells via a pertussis toxin-sensitive mechanism.
    FEBS Lett. 1993 Oct 25;333(1-2):56-60 PMID: 8224171
  18. Regulation of the GTPase cycle of the neuronally expressed Ras-like GTP-binding protein Rab3A.
    J Biol Chem. 1993 Oct 25;268(30):22247-50 PMID: 8226729
  19. Rab proteins and the road maps for intracellular transport.
    Neuron. 1993 Nov;11(5):789-99 PMID: 8240804
  20. Friends and family: the role of the Rab GTPases in vesicular traffic.
    Cell. 1993 Nov 19;75(4):597-601 PMID: 8242735
  21. GP-3, a newly characterized glycoprotein on the inner surface of the zymogen granule membrane, undergoes regulated secretion.
    J Biol Chem. 1994 Mar 25;269(12):9099-104 PMID: 8132647
  22. Membrane association of Rab5 mediated by GDP-dissociation inhibitor and accompanied by GDP/GTP exchange.
    Nature. 1994 Mar 10;368(6467):157-60 PMID: 8139660
  23. Evidence for the involvement of Rab3A in Ca(2+)-dependent exocytosis from adrenal chromaffin cells.
    J Biol Chem. 1994 Apr 8;269(14):10229-34 PMID: 8144603
  24. Pancreatic zymogen granule membrane proteins: molecular details begin to emerge.
    Digestion. 1994;55(4):191-9 PMID: 8063024
  25. A rab protein is required for the assembly of SNARE complexes in the docking of transport vesicles.
    Cell. 1994 Sep 23;78(6):937-48 PMID: 7923363
  26. GTP cleavage by the small GTP-binding protein Rab3A is associated with exocytosis of synaptic vesicles induced by alpha-latrotoxin.
    J Biol Chem. 1994 Oct 7;269(40):24770-6 PMID: 7929154
  27. Actin filament disassembly is a sufficient final trigger for exocytosis in nonexcitable cells.
    J Cell Biol. 1995 Feb;128(4):589-98 PMID: 7860632
  28. The synaptic vesicle cycle: a cascade of protein-protein interactions.
    Nature. 1995 Jun 22;375(6533):645-53 PMID: 7791897
  29. Activation of MAP kinase kinase (MEK) and Ras by cholecystokinin in rat pancreatic acini.
    Am J Physiol. 1995 Jun;268(6 Pt 1):G1060-5 PMID: 7611406
  30. Evidence that the Rab3a-binding protein, rabphilin3a, enhances regulated secretion. Studies in adrenal chromaffin cells.
    J Biol Chem. 1995 Jul 14;270(28):16714-8 PMID: 7622481
  31. Ca(2+)-dependent amylase secretion from SLO-permeabilized rat pancreatic acini requires diffusible cytosolic proteins.
    Am J Physiol. 1995 Nov;269(5 Pt 1):G647-52 PMID: 7491954
  32. rab3D protein is a specific marker for zymogen granules in gastric chief cells of rats and rabbits.
    Gastroenterology. 1996 Mar;110(3):809-20 PMID: 8608891
  33. Distinct functional properties of Rab3A and Rab3B in PC12 neuroendocrine cells.
    J Biol Chem. 1996 Mar 22;271(12):6963-71 PMID: 8636125
  34. Expression of Rab3D in dispersed chief cells from guinea pig stomach.
    Biochim Biophys Acta. 1996 Apr 24;1311(2):111-6 PMID: 8630328
  35. Rab3C is a synaptic vesicle protein that dissociates from synaptic vesicles after stimulation of exocytosis.
    J Biol Chem. 1994 Apr 15;269(15):10971-4 PMID: 8157621
  36. Identification of Rab 5 but not Rab 3A in rat pancreatic zymogen granule membranes.
    Biochem Biophys Res Commun. 1994 Apr 15;200(1):542-8 PMID: 8166729
  37. The GTPase Rab3a negatively controls calcium-dependent exocytosis in neuroendocrine cells.
    EMBO J. 1994 May 1;13(9):2029-37 PMID: 8187757
  38. Rab3D localizes to secretory granules in rat pancreatic acinar cells.
    Eur J Cell Biol. 1996 May;70(1):33-41 PMID: 8738417
  39. Rab3D localizes to zymogen granules in rat pancreatic acini and other exocrine glands.
    Am J Physiol. 1996 Sep;271(3 Pt 1):G531-8 PMID: 8843780
  40. The expression pattern of rab3D in the developing rat exocrine pancreas coincides with the acquisition of regulated exocytosis.
    Eur J Cell Biol. 1996 Oct;71(2):129-36 PMID: 8905289
  41. The vesicle-associated membrane protein family of proteins in rat pancreatic and parotid acinar cells.
    Gastroenterology. 1996 Dec;111(6):1661-9 PMID: 8942747
  42. Distinct cellular locations of the syntaxin family of proteins in rat pancreatic acinar cells.
    Mol Biol Cell. 1996 Dec;7(12):2019-27 PMID: 8970162
  43. Effects of carbachol, cholecystokinin, and insulin on protein phosphorylation in isolated pancreatic acini.
    J Biol Chem. 1982 Sep 10;257(17):10523-8 PMID: 7050109
Article Info
Journal
The Journal of clinical investigation
Abbr.
J Clin Invest
ISSN
0021-9738
DOI
10.1172/JCI119859
Published
1997-12-15
Pages
3044-52
Language
English
Region
United States
NLM ID
7802877
PMCID
PMC508517
Subset
IM
Grants
NIDDK NIH HHS · DK-20572 · United States
NIDDK NIH HHS · DK-34933 · United States
NIDDK NIH HHS · DK-45722 · United States
External Links
PubMed source DOI: 10.1172/JCI119859 PMC Full Text View journal details
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