Inhibition of IKKα by BAY61-3606 Reveals IKKα-Dependent Histone H3 Phosphorylation in Human Cytomegalovirus Infected Cells.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Cytomegalovirus Infections/*metabolism ; Histones/*metabolism ; I-kappa B Kinase/*antagonists & inhibitors ; I-kappa B Kinase/*metabolism ; Niacinamide/*analogs & derivatives ; Phosphorylation/*physiology ; Pyrimidines/*pharmacology; Acetylation/drug effects ; Antigens, Viral/metabolism ; Cells, Cultured ; Cytomegalovirus/drug effects ; Cytomegalovirus/metabolism ; Humans ; Immediate-Early Proteins/metabolism ; NF-kappa B/metabolism ; Niacinamide/pharmacology ; Phosphorylation/drug effects ; Signal Transduction/drug effects ; Viral Proteins/metabolism ; Virus Replication/drug effects

Protein kinase inhibitors can be used as tools to identify proteins and pathways required for virus replication. Using virus replication assays and western blotting we found that the widely used protein kinase inhibitor BAY61-3606 inhibits replication of human cytomegalovirus (HCMV) strain AD169 and the accumulation of HCMV immediate-early proteins in AD169 infected cells, but has no effect on replication of HCMV strain Merlin. Using in vitro kinase assays we found that BAY61-3606 is a potent inhibitor of the cellular kinase IKKα. Infection of cells treated with siRNA targeting IKKα indicated IKKα was required for efficient AD169 replication and immediate-early protein production. We hypothesized that IKKα was required for AD169 immediate-early protein production as part of the canonical NF-κB signaling pathway. However, although BAY61-3606 inhibited phosphorylation of the IKKα substrate IκBα, we found no canonical or non-canonical NF-κB signaling in AD169 infected cells. Rather, we observed that treatment of cells with BAY61-3606 or siRNA targeting IKKα decreased phosphorylation of histone H3 at serine 10 (H3S10p) in western blotting assays. Furthermore, we found treatment of cells with BAY61-3606, but not siRNA targeting IKKα, inhibited the accumulation of histone H3 acetylation (H3K9ac, H3K18ac and H3K27ac) and tri-methylation (H3K27me3 and H3K36me3) modifications. Therefore, the requirement for IKKα in HCMV replication was strain-dependent and during replication of an HCMV strain requiring IKKα, IKKα-dependent H3S10 phosphorylation was associated with efficient HCMV replication and immediate-early protein production. Plus, inhibition of HCMV replication by BAY61-3606 is associated with acetylation and tri-methylation modifications of histone H3 that do not involve IKKα.

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0 (2-(7-(3,4-dimethoxyphenyl)imidazo(1,2-c)pyrimidin-5-ylamino)nicotinamide)
0 (Antigens, Viral)
0 (Histones)
0 (Immediate-Early Proteins)
0 (NF-kappa B)
0 (Pyrimidines)
0 (Viral Proteins)
0 (immediate-early proteins, cytomegalovirus)
25X51I8RD4 (Niacinamide)
EC 2.7.11.10 (I-kappa B Kinase)

Date Created: 20160302 Date Completed: 20160718 Latest Revision: 20190222

20231215

PMC4773098

10.1371/journal.pone.0150339

26930276