title: The spatio-temporal dynamics of PKA activity profile during mitosis and its correlation to chromosome segregation.
abstract: 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.
authors.affiliation: a Laboratoire de Régulation des Signaux de division; EA4479; Université Lille1; Université Lille Nord de France; Villeneuve d'Ascq, France Institut Fédératif de Recherche (IFR)147; Site de Recherche Intégré en Cancérologie (SIRIC) ONCOLILLE.
authors.forename: Pauline
authors.initials: P
authors.lastname: Vandame
authors.forename: Corentin
authors.initials: C
authors.lastname: Spriet
authors.forename: Dave
authors.initials: D
authors.lastname: Trinel
authors.forename: Armance
authors.initials: A
authors.lastname: Gelaude
authors.forename: Katia
authors.initials: K
authors.lastname: Caillau
authors.forename: Coralie
authors.initials: C
authors.lastname: Bompard
authors.forename: Emanuele
authors.initials: E
authors.lastname: Biondi
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Bodart
mesh_terms: D015374:Biosensing Techniques; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology; D020090:Chromosome Segregation / Q000235:genetics / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D008938:Mitosis / Q000235:genetics / Q000502:physiology*
chemical_list: D017868:Cyclic AMP-Dependent Protein Kinases
keywords: Cell cycle; FRET based biosensor; Mitosis; PKA; chromosome segregation;  