The spatio-temporal dynamics of PKA activity profile during mitosis and its correlation to chromosome segregation

The cyclic adenosine monophosphate dependent kinase protein (PKA) controls a variety of cellular processes including cell cycle regulation. Here, we took advantages of genetically encoded FRET-based biosensors, using an AKAR-derived biosensor to characterize PKA activity during mitosis in living HeL...

Full description

Saved in:
Bibliographic Details
Published in:Cell cycle (Georgetown, Tex.) Tex.), 2014-10, Vol.13 (20), p.3232-3240
Main Authors: Vandame, Pauline, Spriet, Corentin, Trinel, Dave, Gelaude, Armance, Caillau, Katia, Bompard, Coralie, Biondi, Emanuele, Bodart, Jean-François
Format: Article
Language:English
Subjects:
Biosensing Techniques
Cell cycle
Cell Cycle - genetics
Cell Cycle - physiology
Chemical Sciences
chromosome segregation
Chromosome Segregation - genetics
Chromosome Segregation - physiology
Cyclic AMP-Dependent Protein Kinases - genetics
Cyclic AMP-Dependent Protein Kinases - metabolism
FRET based biosensor
HeLa Cells
Humans
Microscopy, Confocal
Mitosis
Mitosis - genetics
Mitosis - physiology
or physical chemistry
PKA
Theoretical and
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The cyclic adenosine monophosphate dependent kinase protein (PKA) controls a variety of cellular processes including cell cycle regulation. Here, we took advantages of genetically encoded FRET-based biosensors, using an AKAR-derived biosensor to characterize PKA activity during mitosis in living HeLa cells using a single-cell approach. We measured PKA activity changes during mitosis. HeLa cells exhibit a substantial increase during mitosis, which ends with telophase. An AKAREV T>A inactive form of the biosensor and H89 inhibitor were used to ascertain for the specificity of the PKA activity measured. On a spatial point of view, high levels of activity near to chromosomal plate during metaphase and anaphase were detected. By using the PKA inhibitor H89, we assessed the role of PKA in the maintenance of a proper division phenotype. While this treatment in our hands did not impaired cell cycle progression in a drastic manner, inhibition of PKA leads to a dramatic increase in chromososme misalignement on the spindle during metaphase that could result in aneuploidies. Our study emphasizes the insights that can be gained with genetically encoded FRET-based biosensors, which enable to overcome the shortcomings of classical methologies and unveil in vivo PKA spatiotemporal profiles in HeLa cells.
ISSN:1538-4101
1551-4005
DOI:10.4161/15384101.2014.950907