Visualizing Smad1/4 signaling response to bone morphogenetic protein-4 activation by FRET biosensors.
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Visualizing Smad1/4 signaling response to bone morphogenetic protein-4 activation by FRET biosensors. / Brune, Kira; Friedrich, Mike; Noskov, Andrey; Harms, Gregory S.
In: Biochim Biophys Acta, Vol. 1773, No. 12, 12, 2007, p. 1759-1773.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Visualizing Smad1/4 signaling response to bone morphogenetic protein-4 activation by FRET biosensors.
AU - Brune, Kira
AU - Friedrich, Mike
AU - Noskov, Andrey
AU - Harms, Gregory S
PY - 2007
Y1 - 2007
N2 - Smad proteins are the major signal transducers for the Transforming Growth Factor superfamily of cytokines and their serine/threonine kinase receptors. Smads mediate the signal from the membrane into the nucleus. Bone Morphogenetic Protein-4 stimulates phosphorylation of Smad1, which interacts with Smad4. This complex translocates into the nucleus and regulates transcription of target genes. Here, we report our development of cellular fluorescence biosensors for direct visualization of Smad signaling in live mammalian cells. Fluorescence resonance energy transfer between cyan and yellow fluorescent proteins fused to the Smad1 and Smad4 proteins was used to unravel the temporal aspects of BMP/Smad signaling. A rate-limiting delay of 2-5 min occurred between BMP activation and Smad1 activity. A similar delay was observed in the Smad1/Smad4 complexation. Further experimentation indicated that the delay is dependent on the MH1 domain and linker of Smad1. These results give new insights into the dynamics of the BMP receptor -Smad1/4 signaling process and provide a new tool for studying Smads.
AB - Smad proteins are the major signal transducers for the Transforming Growth Factor superfamily of cytokines and their serine/threonine kinase receptors. Smads mediate the signal from the membrane into the nucleus. Bone Morphogenetic Protein-4 stimulates phosphorylation of Smad1, which interacts with Smad4. This complex translocates into the nucleus and regulates transcription of target genes. Here, we report our development of cellular fluorescence biosensors for direct visualization of Smad signaling in live mammalian cells. Fluorescence resonance energy transfer between cyan and yellow fluorescent proteins fused to the Smad1 and Smad4 proteins was used to unravel the temporal aspects of BMP/Smad signaling. A rate-limiting delay of 2-5 min occurred between BMP activation and Smad1 activity. A similar delay was observed in the Smad1/Smad4 complexation. Further experimentation indicated that the delay is dependent on the MH1 domain and linker of Smad1. These results give new insights into the dynamics of the BMP receptor -Smad1/4 signaling process and provide a new tool for studying Smads.
KW - Animals
KW - Humans
KW - COS Cells
KW - Cercopithecus aethiops
KW - Kinetics
KW - Amino Acid Sequence
KW - Molecular Sequence Data
KW - Protein Structure, Tertiary
KW - Hela Cells
KW - Signal Transduction
KW - Luminescent Proteins/metabolism
KW - Recombinant Fusion Proteins/metabolism
KW - Cell Nucleus/drug effects/metabolism
KW - Protein Transport/drug effects
KW - Biosensing Techniques/methods
KW - Bone Morphogenetic Protein 4
KW - Bone Morphogenetic Proteins/pharmacology
KW - Fluorescence Resonance Energy Transfer/methods
KW - Smad1 Protein/chemistry/metabolism
KW - Smad4 Protein/metabolism
KW - Transcription, Genetic/drug effects
KW - Animals
KW - Humans
KW - COS Cells
KW - Cercopithecus aethiops
KW - Kinetics
KW - Amino Acid Sequence
KW - Molecular Sequence Data
KW - Protein Structure, Tertiary
KW - Hela Cells
KW - Signal Transduction
KW - Luminescent Proteins/metabolism
KW - Recombinant Fusion Proteins/metabolism
KW - Cell Nucleus/drug effects/metabolism
KW - Protein Transport/drug effects
KW - Biosensing Techniques/methods
KW - Bone Morphogenetic Protein 4
KW - Bone Morphogenetic Proteins/pharmacology
KW - Fluorescence Resonance Energy Transfer/methods
KW - Smad1 Protein/chemistry/metabolism
KW - Smad4 Protein/metabolism
KW - Transcription, Genetic/drug effects
M3 - SCORING: Journal article
VL - 1773
SP - 1759
EP - 1773
JO - Biochim Biophys Acta
JF - Biochim Biophys Acta
SN - 0006-3002
IS - 12
M1 - 12
ER -
