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  Imaging CREB activation in living cells

Friedrich, M. W., Aramuni, G., Mank, M., Mackinnon, J. A. G., & Griesbeck, O. (2010). Imaging CREB activation in living cells. The Journal of Biological Chemistry, 285(30), 23285-23295. doi:10.1074/jbc.M110.124545.

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Friedrich, M. W.1, Author              
Aramuni, G.1, Author              
Mank, M.1, Author              
Mackinnon, J. A. G.1, Author              
Griesbeck, O.1, Author              
Affiliations:
1Research Group: Cellular Dynamics / Griesbeck, MPI of Neurobiology, Max Planck Society, ou_1113560              

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 Abstract: The Ca2+- and cAMP-responsive element-binding protein (CREB) and the related ATF-1 and CREM are stimulus-inducible transcription factors that link certain forms of cellular activity to changes in gene expression. They are attributed to complex integrative activation characteristics, but current biochemical technology does not allow dynamic imaging of CREB activation in single cells. Using fluorescence resonance energy transfer between mutants of green fluorescent protein we here develop a signal-optimized genetically encoded indicator that enables imaging activation of CREB due to phosphorylation of the critical serine 133. The indicator of CREB activation due to phosphorylation (ICAP) was used to investigate the role of the scaffold and anchoring protein AKAP79/150 in regulating signal pathways converging on CREB. We show that disruption of AKAP79/150-mediated protein kinase A anchoring or knock-down of AKAP150 dramatically reduces the ability of protein kinase A to activate CREB. In contrast, AKAP79/150 regulation of CREB via L-type channels may only have minor importance. ICAP allows dynamic and reversible imaging in living cells and may become useful in studying molecular components and cell-type specificity of activity-dependent gene expression.

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Language(s): eng - English
 Dates: 2010-07-23
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 493354
ISI: 000279999900062
DOI: 10.1074/jbc.M110.124545
 Degree: -

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Title: The Journal of Biological Chemistry
  Other : JBC
  Abbreviation : J. Biol. Chem.
Source Genre: Journal
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Publ. Info: Baltimore, etc. : American Society for Biochemistry and Molecular Biology [etc.]
Pages: - Volume / Issue: 285 (30) Sequence Number: - Start / End Page: 23285 - 23295 Identifier: ISSN: 0021-9258
CoNE: https://pure.mpg.de/cone/journals/resource/954925410826_1