Protein kinase D1 regulates mitogen progression.

LILIANA G. BIANCIOTTI; GASTON AMABLE ; OSVALDO REY; MARIA ELISA PICCO; EDUARDO MARTINEZ LEON ; LAURA ANAYA ; RODRIGO JACAMO

Buenos Aires

Congreso; Reunión Conjunta de Sociedades Biomédicas.; 2017

Sociedad Argentina de Investigación Clínica

Protein kinase D1 (PKD1) is a serine/threonine kinasethat is activated by pharmacological agents such asphorbol esters and physiological stimuli including GPCRsstimulation, growth factors and oxidative stress amongothers. During the interphase PKD1 play an importantrole in the regulation of a remarkable variety of biologicalprocesses including signal transduction, cell migration,differentiation, proliferation and the development of differenttypes of cancer. Nevertheless very little is knownabout the role of this kinase during other phases of thecell cycle including cell division.Cell division is a complex process regulated by manymacromolecules that are controlled by kinases and phosphataseswhich in turn are regulated by mitotic signalingpathways that remain poorly defined.Hence, the principal aim of this study was to determinewhether PKD1-mediated signaling contributed to mitosisregulation. We employed cells lines expressingeGFP or eGFP-tagged PKD1, under the control of aninducible promoter, to examine by real-time imaging ofsingle-cells maintained under physiological conditionsthe role of PKD1 during mitosis. This methodology wascomplemented by indirect immunofluorescence of fixedcells, Western blot and flow cytometry (FACS). Usingthese methodologies we obtained the following results:1- Catalytic active PKD1 colocalizes with mitotic spindle,centrosomes and midbodies 2- Over-expression of thekinase retards mitosis progression, specifically lengtheningmetaphase duration by 50% (p