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JOURNAL OF VIROLOGY, July 2001, p. 6022–6032 Vol. 75, No. 13 0022-538X/01/$04.00 0 DOI: 10.1128/JVI.75.13.6022–6032.2001 Copyright © 2001, American Society for Microbiology. All Rights Reserved. Human Cytomegalovirus Up-Regulates the Phosphatidylinositol 3-Kinase (PI3-K) Pathway: Inhibition of PI3-K Activity Inhibits Viral Replication and Virus-Induced Signaling ROBERT A. JOHNSON,1,2† XIN WANG,2 XIU-LI MA,2 SHU-MEI HUONG,2 AND ENG-SHANG HUANG1,2,3,4* Department of Microbiology and Immunology, 1 Lineberger Comprehensive Cancer Center,2 Department of Medicine,3 and Curriculum of Genetics and Molecular Biology,4 University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7295 Received 12 December 2000/Accepted 6 April 2001 Infection of quiescent fibroblasts with human cytomegalovirus (HCMV) was found to cause a rapid activa- tion of cellular phosphatidylinositol 3-kinase (PI3-K). Maximum PI3-K activation occurred from 15 to 30 min postinfection. This activation was transient, and by 2 h postinfection (hpi), PI3-K activity had declined to preinfection levels. However, at 4 hpi, a second tier of PI3-K activation was detected, and PI3-K activity remained elevated relative to that of mock-infected cells for the remainder of infection. The cellular kinases Akt and p70S6K and the transcription factor NF-B were activated in a PI3-K-dependent manner at similar times following HCMV infection. Analysis using UV-irradiated virus indicated that no viral protein synthesis wa