تأثيرمركبات لتيولين، ميدستيورين (PKC-412) و PD173074 على مراحل حياة فيروس Cytomegalovirus (HCMV) المختلفة.
Date
2016-04-25
Authors
عامر عزت محمد دوفش
Amer Izzat Mohammad Doufish
Journal Title
Journal ISSN
Volume Title
Publisher
AL-Quds University
جامعة القدس
جامعة القدس
Abstract
The cytoplasmic assembly compartment (AC) in HCMV-infected human foreskin
fibroblasts (HFF) is a unique juxtanuclear ―bulb‖-like structure. Morphology of the AC is
dependent on the activity of the viral-encoded serine/threonine kinase, pUL97. The
―bulb‖-like structure morphology is also altered when wt-HCMV infected cells are treated
with kinase inhibitors NGIC-I, a kinase C inhibitor, or Staurosporine, a wide range
serine/threonine kinase inhibitor. Infection with a UL97 deletion mutant simulated the
inhibition with NGIC-I or Staurosporine of wt-HCMV infection, resulting in a less
compact and a vacuole-rich AC. In all three cases viral titers were reduced 2-3 logs.
Cellular kinases play central roles in regulation of cell replication and differentiation,
making cellular kinase inhibitors attractive antiviral targets. Different protein kinases were
identified to affect different stages of HCMV life cycle and infectivity, which prompted us
to explore yet not recognized kinase inhibitors for their activity against HCMV infection.
In this study, we employed protein kinase inhibitors Luteolin, Midostaurin (PKC-412) and
PD173074, and investigated their influence on the assembly compartment, early stages of
HCMV infection and viral load. Luteolin at 20 μM did indeed reduce viral load without
any detectable effects on the assembly complex. Midostaurin, PKC-412, did not affect
viral load or the assembly compartment. The most striking result was observed with the
tyrosine kinase inhibitor, PD173074, at 5 μM concentration. Although PD173074 did not
affect early stages of HCMV infection, it reduced viral load and induced a specific
structure of the assembly compartment we referred to as ―vesicles’-rich AC pattern.
Hereby, the AC remained its ―bulb‖-like structure, but was remarkably accompanied withvesicles spread throughout the cytoplasm, which arose at 48 h p.i. and were remarkable by
72 h p.i. and 96 h p.i.. These vesicles stained for markers of the assembly complex; viral
tegument proteins pp28 and pp65 as well as WGA Golgi marker and accumulated densely
along the cytoplasmic membrane of the infected cells. Taken together, our data provide the
first evidence for a role of tyrosine kinase in HCMV assembly.
Description
Keywords
العلوم الصيدلانية , Pharmaceutical Sciences