Protein kinase D isozymes activation and localization during mitosis

Abstract

The protein kinase D (PKD) family consists of three serine/threonine protein kinases involved in the regulation of fundamental biological processes in response to their activation and intracellular redistribution. Although a substantial amount of information is available describing the mechanisms regulating the activation and intracellular distribution of the PKD isozymes during interphase, nothing is known of their activation status, localization and role during mitosis. The results presented in this study indicate that during mitosis, PKD3 and PKD are phosphorylated at Ser731 and Ser744 within their activation loop by a mechanism that requires protein kinase C. Mitosis-associated PKD3 Ser731 and PKD Ser744 phosphorylation is related to the catalytic activation of these kinases as evidenced by in vivo phosphorylation of histone deacetylase 5, a substrate of PKD and PKD3. Activation loop-phosphorylated PKD3 and PKD, as well as PKD2, associate with centrosomes, spindles and midbody suggesting that these activated kinases establish dynamic interactions with the mitotic apparatus. Thus, this study reveals a connection between the PKD isozymes and cell division, suggesting a novel role for this family of serine/threonine kinases.

Protein Kinase D3 Sensitizes RAF Inhibitor RAF265 in Melanoma Cells by Preventing Reactivation of MAPK Signaling

“…S3). It was shown that PRKD3 was phosphorylated at its activation loops during mitosis, suggesting that PRKD3 activity is regulated in a cell-cycle dependent manner (35). Although PRKD3 siRNAs or 0.5 mmol/L RAF265 alone resulted in a reduction of G 0 -G 1 cell numbers, we did not observe cooperative effects in reducing G 0 -G 1 cell numbers (Fig.…”

PKD2 and PKD3 Promote Prostate Cancer Cell Invasion via uPA by Shifting Balance Between NF-κB and HDAC1

“…Three members of the PKD family have been identified to date, namely, PKD1 (PKCm), PKD2 and PKD3 (PKCn) (Wang, 2006). PKD has been implicated in various biological processes, such as cell proliferation (McEneaney et al, 2010;Papazyan et al, 2008;Wang, 2006), survival (Arun et al, 2011;Song et al, 2009), cell migration (Eiseler et al, 2010;Peterburs et al, 2009), differentiation (Jadali and Ghazizadeh, 2010;Kleger et al, 2011), inflammation (Kim et al, 2010b;Park et al, 2009) and vesicle transportation (Chen et al, 2009;Hausser et al, 2005;Lu et al, 2007). The role of PKDs in cancer invasion/metastasis has been increasingly recognized, and, of the three PKD isoforms, PKD1 has been the focus of most studies in this context.…”

PKD3 deficiency causes alterations in microtubule dynamics during the cell cycle

“…In addition, earlier work has identified PKD3 and PKD2 as being associated with microtubule structures during mitosis. 14 In the present study, by analyzing genetically depleted cell lines, we identified PKD3 as a modulator of microtubule dynamics during mitosis.…”

“…To date, very few reports have postulated a PKD-or PKD3-specific in vivo function in the context of the cell cycle. Whereas Chen et al 12 have shown that PKD3 modulated Akt and Erk1/2 activities in various prostate cancer cell lines, thereby promoting cell growth, Papazyan et al 14 have reported that PKD3 and PKD2 are specifically localized to microtubule structures during mitosis. In addition, our analysis revealed that in wt MEFs, PKD3 co-localized with microtubules during mitosis (Fig.…”

“…33 Thus, it is very likely that other as-yet undefined kinases, including PKDs, can participate in the regulation of microtubule dynamics. Nonetheless, no reports have identified such a role for PKDs, particularly PKD3, except for the analysis performed by Papazyan et al 14 That report has revealed a microtubule-associated localization of PKD3 in HEK293 cells but included no functional data. Therefore, future research is needed to identify and characterize PKD3-specific substrates among the large number of microtubule-associated proteins to shed light on the molecular mechanism regulated by PKD3.…”