scholarly journals Cytomegaloviral proteins pUL50 and pUL53 are associated with the nuclear lamina and interact with cellular protein kinase C

2007 ◽  
Vol 88 (10) ◽  
pp. 2642-2650 ◽  
Author(s):  
Jens Milbradt ◽  
Sabrina Auerochs ◽  
Manfred Marschall
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Direct Interaction ◽  
Nuclear Lamina ◽  
Cellular Protein ◽  
Immunofluorescent Staining ◽  
Lamin B ◽  
Kinase C ◽  
Lamin B Receptor

Human cytomegalovirus-encoded pUL50 and pUL53 belong to a group of conserved herpesviral nuclear proteins. This study describes: (i) the co-localization of pUL50 with components of the nuclear lamina such as lamins A/C and lamin B receptor by double immunofluorescent staining, (ii) a strong pUL50-mediated relocalization of pUL53 from a diffuse nuclear pattern towards a nuclear rim localization, (iii) a direct interaction between pUL50 and pUL53, as well as between pUL50 and protein kinase C (PKC), shown by yeast two-hybrid and co-immunoprecipitation analyses, (iv) in vitro phosphorylation of pUL50, which is highly suggestive of PKC activity, and finally (v) partial relocalization of PKC by pUL50/pUL53 from its main cytoplasmic localization to a marked nuclear lamina accumulation. These data suggest a role for pUL50 and pUL53 in the recruitment of PKC, an event that is considered to be important for cytomegalovirus-induced distortion of the nuclear lamina.

scholarly journals Herpes Simplex Virus Type 1 Infection Induces Activation and Recruitment of Protein Kinase C to the Nuclear Membrane and Increased Phosphorylation of Lamin B

2006 ◽  
Vol 80 (1) ◽  
pp. 494-504 ◽  
Author(s):  
Richard Park ◽  
Joel D. Baines
Keyword(s):  
Protein Kinase C ◽  
Herpes Simplex ◽  
Protein Kinase ◽  
Nuclear Membrane ◽  
Nuclear Lamina ◽  
Lamin B ◽  
Kinase C ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT We report that herpes simplex virus type 1 (HSV-1) infection leads to the recruitment of protein kinase C (PKC) to the nuclear rim. In HEp-2 cells, PKC recruitment to the nuclear rim was initiated between 8 h and 12 h postinfection. PKCδ, a proapoptotic kinase, was completely recruited to the nuclear rim upon infection with HSV-1. PKCα was less dramatically relocalized mostly at the nuclear rim upon infection, although some PKCα remained in the cytoplasm. PKCζ-specific immunofluorescence was not significantly relocated to the nuclear rim. The UL34 and UL31 proteins, as well as their association, were each required for PKC recruitment to the nuclear rim. The HSV-1 US3 protein product, a kinase which regulates the phosphorylation state and localization of UL34, was not required for PKC recruitment to the nuclear rim; however, it was required for proper localization along the nuclear rim, as PKC appeared unevenly distributed along the nuclear rim of cells infected with US3 null and kinase-dead mutants. HSV-1 infection induced the phosphorylation of both lamin B and PKC. Elevated lamin B phosphorylation in HSV-1-infected cells was partially reduced by inhibitors of PKC. The data suggest a model in which kinases that normally disassemble the nuclear lamina during apoptosis are recruited to the nuclear membrane through functions requiring UL31 and UL34. We hypothesize that the recruitment of PKC functions to phosphorylate lamin B to help modify the nuclear lamina and promote budding of nucleocapsids at the inner nuclear membrane.

In vitro phosphorylation of lamin B by protein kinase C in friend erythroleukemia. Effect of chemically induced differentiation

1991 ◽  
Vol 15 (5) ◽  
pp. 409-426 ◽  
Author(s):  
A BILLI ◽  
A MATTEUCCI ◽  
V BERTAGNOLO ◽  
M PREVIATI ◽  
F MANZOLI ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Induced Differentiation ◽  
Lamin B ◽  
Kinase C ◽  
Chemically Induced

scholarly journals 14–3-3 Inhibits the Dictyostelium Myosin II Heavy-Chain-specific Protein Kinase C Activity by a Direct Interaction: Identification of the 14–3-3 Binding Domain

1997 ◽  
Vol 8 (10) ◽  
pp. 1889-1899 ◽  
Author(s):  
Meirav Matto-Yelin ◽  
Alastair Aitken ◽  
Shoshana Ravid
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Myosin Ii ◽  
Direct Interaction ◽  
Specific Protein ◽  
Kinase C ◽  
C1 Domain ◽  
Specific Protein Kinase

Myosin II heavy chain (MHC) specific protein kinase C (MHC-PKC), isolated from Dictyostelium discoideum, regulates myosin II assembly and localization in response to the chemoattractant cyclic AMP. Immunoprecipitation of MHC-PKC revealed that it resides as a complex with several proteins. We show herein that one of these proteins is a homologue of the 14–3-3 protein (Dd14–3-3). This protein has recently been implicated in the regulation of intracellular signaling pathways via its interaction with several signaling proteins, such as PKC and Raf-1 kinase. We demonstrate that the mammalian 14–3-3 ζ isoform inhibits the MHC-PKC activity in vitro and that this inhibition is carried out by a direct interaction between the two proteins. Furthermore, we found that the cytosolic MHC-PKC, which is inactive, formed a complex with Dd14–3-3 in the cytosol in a cyclic AMP-dependent manner, whereas the membrane-bound active MHC-PKC was not found in a complex with Dd14–3-3. This suggests that Dd14–3-3 inhibits the MHC-PKC in vivo. We further show that MHC-PKC binds Dd14–3-3 as well as 14–3-3ζ through its C1 domain, and the interaction between these two proteins does not involve a peptide containing phosphoserine as was found for Raf-1 kinase. Our experiments thus show an in vivo function for a member of the 14–3-3 family and demonstrate that MHC-PKC interacts directly with Dd14–3-3 and 14–3-3ζ through its C1 domain both in vitro and in vivo, resulting in the inhibition of the kinase.

scholarly journals Direct interaction between p47phox and protein kinase C: evidence for targeting of protein kinase C by p47phox in neutrophils

1999 ◽  
Vol 344 (3) ◽  
pp. 859-866 ◽  
Author(s):  
Emer P. REEVES ◽  
Lodewijk V. DEKKER ◽  
Louisa V. FORBES ◽  
Frans B. WIENTJES ◽  
Ann GROGAN ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Gel Filtration ◽  
Direct Interaction ◽  
Subcellular Fractionation ◽  
Intact Cells ◽  
Kinase C ◽  
C Terminus ◽  
Time Courses

p47phox is an essential component of the NADPH oxidase, and phosphorylation of p47phox is associated with activation of the enzyme. Here we have used p47phox affinity chromatography to extract a p47phox kinase from neutrophil cytosol. The kinase activity was purified by gel filtration and Mini Q chromatography and shown to be indistinguishable from the catalytic fragments of protein kinase C (PKC)-βI, -βII and -∆. The C-terminus of p47phox represented the site of interaction with PKC. Co-immunoprecipitation experiments revealed that the interaction between PKC isotypes and p47phox takes place in intact cells. However PKC-β and -∆ showed different time courses of co-immunoprecipitation, suggesting that the interactions may serve different functions for the various PKC isotypes. Using cells lacking p47phox, we investigated the functional relevance of the interaction between PKC and p47phox. Subcellular fractionation revealed an abnormal recruitment of PKC-βI and -βII, but not PKC-∆, to particulate fractions in p47phox-deficient cells. Phosphorylation of cytosolic proteins was generally increased in stimulated p47phox-deficient neutrophils as compared with normal neutrophils. Furthermore, the cytoskeletal protein coronin was not phosphorylated upon stimulation of p47phox-deficient neutrophils. These findings were confirmed in an in vitro-reconstituted system using rat brain cytosol in which addition of p47phox affected phosphorylation by PKC/PKM (PKM is the catalytic fragment of PKC). These results indicate that p47phox can act as a regulator of PKC in neutrophils.

scholarly journals α-Tocopherol specifically inactivates cellular protein kinase C α by changing its phosphorylation state

1998 ◽  
Vol 334 (1) ◽  
pp. 243-249 ◽  
Author(s):  
Roberta RICCIARELLI ◽  
Andrea TASINATO ◽  
Sophie CLÉMENT ◽  
Nesrin K. ÖZER ◽  
Daniel BOSCOBOINIK ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Smooth Muscle Cells ◽  
Protein Phosphatase ◽  
Antioxidant Properties ◽  
Muscle Cells ◽  
Cellular Protein ◽  
Kinase C

The mechanism of protein kinase C (PKC) regulation by α-tocopherol has been investigated in smooth-muscle cells. Treatment of rat aortic A7r5 smooth-muscle cells with α-tocopherol resulted in a time- and dose-dependent inhibition of PKC. The inhibition was not related to a direct interaction of α-tocopherol with the enzyme nor with a diminution of its expression. Western analysis demonstrated the presence of PKCα, β, δ, ε, ζ and µ isoforms in these cells. Autophosphorylation and kinase activities of the different isoforms have shown that only PKCα was inhibited by α-tocopherol. The inhibitory effects were not mimicked by β-tocopherol, an analogue of α-tocopherol with similar antioxidant properties. The inhibition of PKCα by α-tocopherol has been found to be associated with its dephosphorylation. Moreover the finding of an activation of protein phosphatase type 2A in vitro by α-tocopherol suggests that this enzyme might be responsible for the observed dephosphorylation and subsequent deactivation of PKCα. It is therefore proposed that PKCα inhibition by α-tocopherol is linked to the activation of a protein phosphatase, which in turn dephosphorylates PKCα and inhibits its activity.

In vitro and in vivo suppression of growth of rat liver epithelial tumor cells by antisense oligonucleotide against protein kinase C-alpha

2000 ◽  
Vol 33 (4) ◽  
pp. 601-608 ◽  
Author(s):  
Shwu-Bin Lin ◽  
Li-Ching Wu ◽  
Siao-Ling Huang ◽  
Hui-Lun Hsu ◽  
Sung-Hwa Hsieh ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Tumor Cells ◽  
Rat Liver ◽  
Antisense Oligonucleotide ◽  
Epithelial Tumor ◽  
Kinase C ◽  
Epithelial Tumor Cells
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