scholarly journals Varicella-Zoster Virus (VZV) ORF65 Virion Protein Is Dispensable for Replication in Cell Culture and Is Phosphorylated by Casein Kinase II, but Not by the VZV Protein Kinases

Virology ◽  
2001 ◽  
Vol 280 (1) ◽  
pp. 62-71 ◽  
Author(s):  
Jeffrey I. Cohen ◽  
Hitoshi Sato ◽  
Shamala Srinivas ◽  
Kristen Lekstrom
Keyword(s):  
Cell Culture ◽  
Varicella Zoster Virus ◽  
Protein Kinases ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Virion Protein ◽  
Varicella Zoster

scholarly journals Varicella-Zoster Virus Fc Receptor Component gI Is Phosphorylated on Its Endodomain by a Cyclin-Dependent Kinase

1999 ◽  
Vol 73 (2) ◽  
pp. 1320-1330 ◽  
Author(s):  
Ming Ye ◽  
Karen M. Duus ◽  
Junmin Peng ◽  
David H. Price ◽  
Charles Grose
Keyword(s):  
Fusion Protein ◽  
Varicella Zoster Virus ◽  
Phosphorylation Site ◽  
Cytoplasmic Tail ◽  
Casein Kinase Ii ◽  
Fc Receptor ◽  
Casein Kinase ◽  
Cyclin Dependent Kinase ◽  
Varicella Zoster

Varicella-zoster virus (VZV) glycoprotein gI is a type 1 transmembrane glycoprotein which is one component of the heterodimeric gE:gI Fc receptor complex. Like VZV gE, VZV gI was phosphorylated in both VZV-infected cells and gI-transfected cells. Preliminary studies demonstrated that a serine 343-proline 344 sequence located within the gI cytoplasmic tail was the most likely phosphorylation site. To determine which protein kinase catalyzed the gI phosphorylation event, we constructed a fusion protein, consisting of glutathione-S-transferase (GST) and the gI cytoplasmic tail, called GST-gI-wt. When this fusion protein was used as a substrate for gI phosphorylation in vitro, the results demonstrated that GST-gI-wt fusion protein was phosphorylated by a representative cyclin-dependent kinase (CDK) called P-TEFb, a homologue of CDK1 (cdc2). When serine 343 within the serine-proline phosphorylation site was replaced with an alanine residue, the level of phosphorylation of the gI fusion protein was greatly reduced. Subsequent experiments with individually immunoprecipitated mammalian CDKs revealed that the VZV gI fusion protein was phosphorylated best by CDK1, to a lesser degree by CDK2, and not at all by CDK6. Transient-transfection assays carried out in the presence of the specific CDK inhibitor roscovitine strongly supported the prior results by demonstrating a marked decrease in gI phosphorylation while gI protein expression was unaffected. Finally, the possibility that VZV gI contained a CDK phosphorylation site in its endodomain was of further interest because its partner, gE, contains a casein kinase II phosphorylation site in its endodomain; prior studies have established that CDK1 can phosphorylate casein kinase II.

scholarly journals Phosphorylation of the Nuclear Form of Varicella-Zoster Virus Immediate-Early Protein 63 by Casein Kinase II at Serine 186

2009 ◽  
Vol 83 (23) ◽  
pp. 12094-12100 ◽  
Author(s):  
Niklaus H. Mueller ◽  
Laurie L. Graf ◽  
David Orlicky ◽  
Don Gilden ◽  
Randall J. Cohrs
Keyword(s):  
Monoclonal Antibody ◽  
Varicella Zoster Virus ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Antibody Binding ◽  
Immediate Early ◽  
Productive Infection ◽  
Varicella Zoster ◽  
Recombinant Monoclonal Antibody ◽  
Early Protein

ABSTRACT Varicella-zoster virus (VZV) open reading frame (ORF) 63 is abundantly transcribed in latently infected human ganglia and encodes a 278-amino-acid protein, IE63, with immediate-early kinetics. IE63 is expressed in the cytoplasm of neurons during VZV latency and in both the cytoplasm and the nucleus during productive infection; however, the mechanism(s) involved in IE63 nuclear import and retention has remained unclear. We constructed and identified a recombinant monoclonal antibody to detect a posttranslationally modified form of IE63. Analysis of a series of IE63 truncation and substitution mutants showed that amino acids 186 to 195 are required for antibody binding. Synthetic peptides corresponding to this region identified IE63 S186 as a target for casein kinase II phosphorylation. In addition, acidic charges supplied by E194 and E195 were required for antibody binding. Immunofluorescence analysis of VZV-infected MeWo cells using the recombinant monoclonal antibody detected IE63 exclusively in the nuclei of infected cells, indicating that casein kinase II phosphorylation of S186 occurs in the nucleus and possibly identifying an initial molecular event operative in VZV reactivation.

Comparison of varicella-zoster virus ORF47 protein kinase and casein kinase II and their substrates

2003 ◽  
Vol 70 (S1) ◽  
pp. S95-S102 ◽  
Author(s):  
T.K. Kenyon ◽  
Elizabeth Homan ◽  
J. Storlie ◽  
Minako Ikoma ◽  
Charles Grose
Keyword(s):  
Protein Kinase ◽  
Varicella Zoster Virus ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Varicella Zoster

scholarly journals Identifying and characterizing casein kinase II in human platelets

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3517-3523 ◽  
Author(s):  
CH Hoyt ◽  
CJ Oh ◽  
JB Beekman ◽  
DW Litchfield ◽  
KM Lerea
Keyword(s):  
Protein Kinase ◽  
Protein Kinases ◽  
Casein Kinase Ii ◽  
Human Platelets ◽  
Casein Kinase ◽  
Immunogold Electron Microscopy ◽  
Beta Subunits ◽  
Specific Substrate ◽  
Substrate Peptide ◽  
Independent Protein

Abstract We have recently shown that inhibition of protein phosphatases in platelets causes increases in protein phosphorylations with a concomitant inhibition of platelet responses. The burst in protein phosphorylation appears to be catalyzed by messenger-independent protein kinases. The aim of the present study was to characterize the presence of broad families of protein kinases found in platelets. Lysates of control and thrombin-stimulated platelets were prepared, and proteins were separated on MONO Q fast protein liquid chromatography. In addition to the presence of histone protein kinase and tyrosine kinase activities, human platelets contain casein kinase II (CKII) activity as assessed by phosphorylation of a specific substrate peptide. Western blot analysis and immunogold electron microscopy studies further showed the presence of alpha-, alpha'-, and beta- subunits of CKII. The enzyme appears to be distributed throughout the cytosol and not secreted after thrombin treatment. Immunoprecipitation studies suggest that at least some of the holoenzymes exist as an alpha alpha' beta 2 complex. Although no activation of the enzyme was detected after thrombin treatment, our results show that CKII is a major messenger-independent protein kinase in platelets.

scholarly journals DNA repair protein O6-alkylguanine-DNA alkyltransferase is phosphorylated by two distinct and novel protein kinases in human brain tumour cells

2000 ◽  
Vol 351 (2) ◽  
pp. 393-402 ◽  
Author(s):  
Srinivas R. S. MULLAPUDI ◽  
Francis ALI-OSMAN ◽  
Jiang SHOU ◽  
Kalkunte S. SRIVENUGOPAL
Keyword(s):  
Dna Repair ◽  
Protein Kinase ◽  
Tyrosine Kinase ◽  
Protein Kinases ◽  
Casein Kinase Ii ◽  
Casein Kinase ◽  
Biochemical Modulation ◽  
Cell Extracts ◽  
Dna Alkyltransferase

We showed recently that human O6-alkylguanine-DNA alkyltransferase (AGT), an important target for improving cancer chemotherapy, is a phosphoprotein and that phosphorylation inhibits its activity [Srivenugopal, Mullapudi, Shou, Hazra and Ali-Osman (2000) Cancer Res. 60, 282–287]. In the present study we characterized the cellular kinases that phosphorylate AGT in the human medulloblastoma cell line HBT228. Crude cell extracts used Mg2+ more efficiently than Mn2+ for phosphorylating human recombinant AGT (rAGT) protein. Both [γ-32P]ATP and [γ-32P]GTP served as phosphate donors, with the former being twice as efficient. Specific components known to activate protein kinase A, protein kinase C and calmodulin-dependent kinases did not stimulate the phosphorylation of rAGT. Phosphoaminoacid analysis after reaction in vitro with ATP or GTP showed that AGT was modified at the same amino acids (serine, threonine and tyrosine) as in intact HBT228 cells. Although some of these properties pointed to casein kinase II as a candidate enzyme, known inhibitors and activators of casein kinase II did not affect rAGT phosphorylation. Fractionation of the cell extracts on poly(Glu/Tyr)-Sepharose resulted in the adsorption of an AGT kinase that modified the tyrosine residues and the exclusion of a fraction that phosphorylated AGT on serine and threonine residues. In-gel kinase assays after SDS/PAGE and non-denaturing PAGE revealed the presence of two AGT kinases of 75 and 130kDa in HBT228 cells. The partly purified tyrosine kinase, identified as the 130kDa enzyme by the same assays, was strongly inhibited by tyrphostin 25 but not by genestein. The tyrosine kinase used ATP or GTP to phosphorylate the AGT protein; this reaction inhibited the DNA repair activity of AGT. Evidence that the kinases might physically associate with AGT in cells was also provided. These results demonstrate that two novel cellular protein kinases, a tyrosine kinase and a serine/threonine kinase, both capable of using GTP as a donor, phosphorylate the AGT protein and affect its function. The new kinases might serve as potential targets for strengthening the biochemical modulation of AGT in human tumours.

Genotyping of varicella-zoster virus strains after serial passages in cell culture

2007 ◽  
Vol 145 (1) ◽  
pp. 80-83 ◽  
Author(s):  
Andreas Sauerbrei ◽  
Anja Philipps ◽  
Roland Zell ◽  
Peter Wutzler
Keyword(s):  
Cell Culture ◽  
Varicella Zoster Virus ◽  
Varicella Zoster ◽  
Virus Strains
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