PROTEIN KINASE D1 REGULATES MITOSIS PROGRESSION.

E. MARTÍNEZ LEÓN; R. JACAMO; G. AMABLE; M. E. PICCO; L. ANAYA; L. BIANCIOTTI; O. REY

Buenos Aires

Congreso; Reunión Conjunta de Sociedades de Biociencias; 2017

Sociedad Argentina de Investigación Clínica

&amp;lt;!-- /* Font Definitions */ @font-face{font-family:"Cambria Math";panose-1:2 4 5 3 5 4 6 3 2 4;mso-font-charset:0;mso-generic-font-family:roman;mso-font-pitch:variable;mso-font-signature:-536870145 1107305727 0 0 415 0;}@font-face{font-family:Calibri;panose-1:2 15 5 2 2 2 4 3 2 4;mso-font-charset:0;mso-generic-font-family:swiss;mso-font-pitch:variable;mso-font-signature:-536859905 -1073732485 9 0 511 0;} /* Style Definitions */ p.MsoNormal, li.MsoNormal, div.MsoNormal{mso-style-unhide:no;mso-style-qformat:yes;mso-style-parent:"";margin:0cm;margin-bottom:.0001pt;mso-pagination:widow-orphan;font-size:12.0pt;font-family:"Calibri",sans-serif;mso-ascii-font-family:Calibri;mso-ascii-theme-font:minor-latin;mso-fareast-font-family:Calibri;mso-fareast-theme-font:minor-latin;mso-hansi-font-family:Calibri;mso-hansi-theme-font:minor-latin;mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi;mso-ansi-language:ES-TRAD;mso-fareast-language:EN-US;}.MsoChpDefault{mso-style-type:export-only;mso-default-props:yes;font-family:"Calibri",sans-serif;mso-ascii-font-family:Calibri;mso-ascii-theme-font:minor-latin;mso-fareast-font-family:Calibri;mso-fareast-theme-font:minor-latin;mso-hansi-font-family:Calibri;mso-hansi-theme-font:minor-latin;mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi;mso-ansi-language:ES-TRAD;mso-fareast-language:EN-US;}@page WordSection1{size:612.0pt 792.0pt;margin:70.85pt 3.0cm 70.85pt 3.0cm;mso-header-margin:36.0pt;mso-footer-margin:36.0pt;mso-paper-source:0;}div.WordSection1{page:WordSection1;}--&amp;gt;Protein kinase D1 (PKD1) is a serine/threonine kinase that is activated by pharmacological agents such as phorbol esters and physiological stimuliincluding GPCRs stimulation, growth factors and oxidative stress among others. During the interphase PKD1 play an important role in the regulation of a remarkable variety of biological processes including signal transduction, cell migration, differentiation, proliferation and the development of different types of cancer. Nevertheless very little is knownabout the role of this kinase during other phases of the cell cycle includingcell division.Cell division is a complex process regulated by many macromolecules that are controlled by kinases and phosphatases which in turn are regulated by mitotic signaling pathwaysthat remain poorly defined.Hence, the principal aim of this study was to determine whetherPKD1-mediated signaling contributed to mitosis regulation. We employed cellslines expressingeGFP or eGFP-tagged PKD1, under thecontrol of an inducible promoter, to examine byreal-time imaging of single-cells maintained under physiological conditions therole of PKD1 during mitosis. This methodology was complemented by indirect immunofluorescenceof fixed cells, Western blot and flow cytometry (FACS).  Using these methodologies we obtained thefollowing results: 1- Catalytic active PKD1 colocalizes with mitotic spindle, centrosomesand midbodies 2- Over-expression of the kinase retards mitosis progression, specificallylengthening metaphase duration by 50% (p<0.001) 3- Inhibition of PKD1activity prior to entry into mitosis stops cell cycle progression (Coefficientof variation (CV)<8, 0) 4- Cells that were incubated in the presence of PKD1specific inhibitors, after entrance into the mitotic phase, do not progressthrough the cycle preventing the completion of mitosis (CV<8.0). These data,therefore, indicates that PKD1 regulates the progression of cellular mitosis.