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Title: Human Cytomegalovirus UL97 Kinase Activity Is Required for the Hyperphosphorylation of Retinoblastoma Protein and Inhibits the Formation of Nuclear Aggresomes

Abstract

Cells infected with human cytomegalovirus in the absence of UL97 kinase activity produce large nuclear aggregates that sequester considerable quantities of viral proteins. A transient expression assay suggested that pp71 and IE1 were also involved in this process, and this suggestion was significant, since both proteins have been reported to interact with components of promyelocytic leukemia (PML) bodies (ND10) and also interact functionally with retinoblastoma pocket proteins (RB). PML bodies have been linked to the formation of nuclear aggresomes, and colocalization studies suggested that viral proteins were recruited to these structures and that UL97 kinase activity inhibited their formation. Proteins associated with PML bodies were examined by Western blot analysis, and pUL97 appeared to specifically affect the phosphorylation of RB in a kinasedependent manner. Three consensus RB binding motifs were identified in the UL97 kinase, and recombinant viruses were constructed in which each was mutated to assess a potential role in the phosphorylation of RB and the inhibition of nuclear aggresome formation. The mutation of either the conserved LxCxE RB binding moti for the lysine required for kinase activity impaired the ability of the virus to stabilize and phosphorylate RB. We concluded from these studies that both UL97 kinase activitymore » and the LxCxE RB binding motif are required for the phosphorylation and stabilization of RB in infected cells and that this effect can be antagonized by the antiviral drug maribavir. These data also suggest a potential link between RB function and the formation of aggresomes.« less

Authors:
; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
972976
Report Number(s):
PNNL-SA-63189
Journal ID: ISSN 0022-538X; JOVIAM; KC0304000; TRN: US201005%%479
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Journal of Virology, 82(10):5054-5067
Additional Journal Information:
Journal Volume: 82; Journal Issue: 10; Journal ID: ISSN 0022-538X
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; LEUKEMIA; LYSINE; MUTATIONS; PHOSPHORYLATION; PHOSPHOTRANSFERASES; PROTEINS; RUTHERFORD BACKSCATTERING SPECTROSCOPY; STABILIZATION; TRANSIENTS; VIRUSES

Citation Formats

Prichard, Mark N., Sztul, Elizabeth, Daily, Shannon L., Perry, Amie L., Frederick, Samuel L., Gill, Rachel B., Hartline, Caroll B., Streblow, Daniel N., Varnum, Susan M., Smith, Richard D., and Kern, Earl R. Human Cytomegalovirus UL97 Kinase Activity Is Required for the Hyperphosphorylation of Retinoblastoma Protein and Inhibits the Formation of Nuclear Aggresomes. United States: N. p., 2008. Web. doi:10.1128/JVI.02174-07.
Prichard, Mark N., Sztul, Elizabeth, Daily, Shannon L., Perry, Amie L., Frederick, Samuel L., Gill, Rachel B., Hartline, Caroll B., Streblow, Daniel N., Varnum, Susan M., Smith, Richard D., & Kern, Earl R. Human Cytomegalovirus UL97 Kinase Activity Is Required for the Hyperphosphorylation of Retinoblastoma Protein and Inhibits the Formation of Nuclear Aggresomes. United States. doi:10.1128/JVI.02174-07.
Prichard, Mark N., Sztul, Elizabeth, Daily, Shannon L., Perry, Amie L., Frederick, Samuel L., Gill, Rachel B., Hartline, Caroll B., Streblow, Daniel N., Varnum, Susan M., Smith, Richard D., and Kern, Earl R. Thu . "Human Cytomegalovirus UL97 Kinase Activity Is Required for the Hyperphosphorylation of Retinoblastoma Protein and Inhibits the Formation of Nuclear Aggresomes". United States. doi:10.1128/JVI.02174-07.
@article{osti_972976,
title = {Human Cytomegalovirus UL97 Kinase Activity Is Required for the Hyperphosphorylation of Retinoblastoma Protein and Inhibits the Formation of Nuclear Aggresomes},
author = {Prichard, Mark N. and Sztul, Elizabeth and Daily, Shannon L. and Perry, Amie L. and Frederick, Samuel L. and Gill, Rachel B. and Hartline, Caroll B. and Streblow, Daniel N. and Varnum, Susan M. and Smith, Richard D. and Kern, Earl R.},
abstractNote = {Cells infected with human cytomegalovirus in the absence of UL97 kinase activity produce large nuclear aggregates that sequester considerable quantities of viral proteins. A transient expression assay suggested that pp71 and IE1 were also involved in this process, and this suggestion was significant, since both proteins have been reported to interact with components of promyelocytic leukemia (PML) bodies (ND10) and also interact functionally with retinoblastoma pocket proteins (RB). PML bodies have been linked to the formation of nuclear aggresomes, and colocalization studies suggested that viral proteins were recruited to these structures and that UL97 kinase activity inhibited their formation. Proteins associated with PML bodies were examined by Western blot analysis, and pUL97 appeared to specifically affect the phosphorylation of RB in a kinasedependent manner. Three consensus RB binding motifs were identified in the UL97 kinase, and recombinant viruses were constructed in which each was mutated to assess a potential role in the phosphorylation of RB and the inhibition of nuclear aggresome formation. The mutation of either the conserved LxCxE RB binding moti for the lysine required for kinase activity impaired the ability of the virus to stabilize and phosphorylate RB. We concluded from these studies that both UL97 kinase activity and the LxCxE RB binding motif are required for the phosphorylation and stabilization of RB in infected cells and that this effect can be antagonized by the antiviral drug maribavir. These data also suggest a potential link between RB function and the formation of aggresomes.},
doi = {10.1128/JVI.02174-07},
journal = {Journal of Virology, 82(10):5054-5067},
issn = {0022-538X},
number = 10,
volume = 82,
place = {United States},
year = {2008},
month = {5}
}