Title page for ETD etd-07082008-083329


Type of Document Master's Thesis
Author Lee, HyunCheol
URN etd-07082008-083329
Title Comparative Roles of Herpes Simplex Virus Type 1 (HSV-1) Glycoproteins in Cytoplasmic Virion Egress
Degree Master of Science (M.S.)
Department Pathobiological Sciences (Veterinary Medical Sciences)
Advisory Committee
Advisor Name Title
Konstantin Gus Kousoulas Committee Chair
Marlene S. Orandle Committee Member
Masami Yoshimura Committee Member
Keywords
  • cytoplasmic envelopment
  • glycoproteins
  • HSV-1
Date of Defense 2008-06-27
Availability unrestricted
Abstract
HSV-1 acquires its final envelope by budding into cytoplasmic vesicles thought to be

derived from Trans-Golgi Network (TGN) membranes. This process is facilitated by

interactions among the carboxyl termini of viral glycoproteins and tegument proteins. To

investigate the relative importance of different viral glycoproteins in cytoplasmic virion

morphogenesis, a set of recombinant viruses were constructed silencing expression of

the glycoprotein E (ΔgE), the carboxyl terminus of glycoprotein D (gDΔcp), and the

membrane protein UL20 (ΔUL20). In addition, recombinant viruses were constructed

having the ΔgE+gDΔcp, and the ΔgE+ΔgM (glycoprotein M) deletions. These

recombinant viruses were constructed using the double-red, site-directed mutagenesis

system implemented on the HSV-1 genome cloned into a bacterial artificial

chromosome (bac). The ΔgE, ΔgE+ΔgM, and gDΔcp viruses produced viral plaques

that were approximately 50% smaller to those produced by the wild-type virus HSV-1(F

strain). The gDΔcp+ΔgE recombinant virus produced viral plaques that were on the

average 50% smaller to those produced by either the ΔgE, ΔgE+ΔgM, or the gDΔcp

viruses. However, these viral plaques were substantially larger than those produced by

previously constructed UL20-null or gK-null viruses. Kinetics of viral replication revealed

that all recombinant viruses appeared to produce similar viral titers at late times post

infection. However, both the gDΔcp and the ΔgE+gDΔcp viruses appeared to replicate

slower than the wild-type virus or the ΔgE and ΔgE+ΔgM viruses. Electron microscopy

revealed that all viruses regardless of their different gene mutations produced

enveloped virions that were secreted outside, with no apparent accumulation of

unenveloped capsids in the cytoplasm of infected cells. These results suggest that the

gD and gE carboxyl termini, either alone or in a redundant manner, are not essential in

cytoplasmic virion envelopment and egress from infected cells. Furthermore, the results

show that gK/UL20 complex serves preeminent roles among all viral glycoproteins in

cytoplasmic virion morphogenesis and egress.

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