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In situ regulation of cell-cell communication by the cAMP-dependent protein kinase and protein kinase C

  • Angela J. Godwin
  • Lora M. Green
  • Michael P. Walsh
  • John R. McDonald
  • Donal A. Walsh
  • William H. Fletcher
Chapter
Part of the Developments in Molecular and Cellular Biochemistry book series (DMCB, volume 11)

Abstract

The effects of cAMP-dependent protein kinase A and protein kinase C on cell-cell communication have been exam ined in primary ovarian granulosa cells microinjected with purified components of these two regulatory cascades. These cells possess connexin43 (α l)-type gap junctions, and are well-coupled electrotonically and as judged by the cell-to-cell transfer of fluorescent dye. Within 2–3 min after injection of the protein kinase A inhibitor (PKI) communication was sharply reduced or ceased, but resumed in about 3 min with the injection of the protein kinase A catalytic subunit. A similar resumption also occurred in PKI-injected cells after exposure to follicle stimulating hormone. Microinjection of the protein kinase C inhibitor protein caused a transient cessation of communication that spontaneously returned within 15–20 min. Treatment of cells with activators of protein kinase C, TPA or OAG for 60 min caused a significant reduction in communication that could be restored within 2–5 min by the subsequent injection of either the protein kinase C inhibitor or the protein kinase A catalytic subunit. With a longer exposure to either protein kinase C activator communication could not be restored and this appeared to be related to the absence of aggregates of connexin43 in membrane as detected immunologically. In cells injected with alkaline phosphatase communication stopped but returned either spontaneously within 20 min or within 2–3 min of injecting the cell with either the protein kinase A catalytic subunit or with protein kinase C. When untreated cells were injected with protein kinase C communication diminished or ceased within 5 min. Collectively these results demonstrate that cell-cell communication is regulated by both protein kinase A and C, but in a complex interrelated manner, quite likely by multiple phosphorylation of proteins within or regulating connexin-43 containing gap junctions. (Mol Cell Biochem 127/128: 293–307, 1993)

Key words

cell-cell communication cyclic AMP-dependent protein kinase protein kinase C 

Abbreviations

C

catalytic subunit of protein kinase A

CKI

protein kinase C inhibitor protein

Cx

connexin protein

dbcAMP

N6,2′-O-dibutyryladenosine 3′:5′-cyclic monophosphate

OAG

1-oleoyl-2-acetyl-sn-glycerol

protein kinase A

cAMP-dependent protein kinase

protein kinase C

Ca2+-sensitive phospholipid-dependent protein kinase

PKI

protein kinase A inhibitor protein

R

regulatory subunit of protein kinase A

TPA

12-O-tetradecanoylphorbol-13-acetate

8Br-cAMP

8-bromoadenosine 3′:5′ cyclic monophosphate

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Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Angela J. Godwin
    • 1
  • Lora M. Green
    • 2
  • Michael P. Walsh
    • 3
  • John R. McDonald
    • 4
  • Donal A. Walsh
    • 5
  • William H. Fletcher
    • 1
  1. 1.Department of AnatomyLoma Linda University and Molecular Cytology Research, Veterans Administration Medical CenterLoma LindaUSA
  2. 2.Department of MicrobiologyLoma Linda University and Immunology Research, Veterans Administration Medical CenterLoma LindaUSA
  3. 3.Department of Medical BiochemistryUniversity of CalgaryCanada
  4. 4.Synergen Inc.BoulderUSA
  5. 5.Department of Biological ChemistryUniversity of CaliforniaDavisUSA

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