Areas of Interest
HSV-1 and -2 are large DNA virus which are responsible for oral
and genital infections, sight-threatening ocular infections and
brain infections which can be life threatening. Our laboratory
uses genetic, biochemical, biophysical, structural and molecular
approaches to elucidate mechanisms of synthesis, maturation and
cleavage and packaging of viral genomes. One of our goals is to
identify antiviral targets which can be exploited to develop
strategies for controlling viral infections. Projects in the lab
are diverse and include virus-host interactions important for
the formation of replication compartments within the nucleus of
infected cells; structure function analysis of two viral
helicases; structure-function analysis of the zinc binding
domain of the HSV-1 primase; analysis of the role of
recombination in viral DNA replication and genome maturation and
the analysis of the machinery involved in cleavage and packaging
of viral genomes into preformed procapsids. Important recent
discoveries include:
- The identification and characterization of a novel viral
recombinase.
- The realization that host cell recombination and repair
proteins are involved in HSV DNA synthesis.
- The discovery that host chaperones are rearranged in
infected cells into foci which act as quality control
mechanisms for the production of properly folded proteins.
- The finding that the initiation helicase-origin binding
protein UL9 is degraded during infection in a ubiquitin
mediated pathway.
- The further characterization of the interdependence of
the helicase and primase subunits with each other.
Lab Rotation Projects
The role of the cellular DNA damage response in the Herpes
Simplex Virus Life Cycle.
It is becoming clear that viruses have evolved elaborate
interactions with the cellular repair, recombination and
checkpoint machinery in order to create an environment conducive
to their own replication. The host cell’s DNA damage machinery
is alert for perturbations in DNA which could lead to genetic
instability. After infection, some of this machinery is
inactivated by the virus in attempt to remove obstacles to
productive infection; however, other components are utilized by
the virus to promote viral DNA replication. In this project,
aspects of this fascinating cat and mouse game will be examined.
Cleavage and Packaging of Herpes Simplex Virus genomes.
Capsid assembly and genome encapsidation are critical aspects in
the life cycle of any virus. Our goal is to gain a better
understanding of the processes by which head to tail
concatemeric DNA molecules are taken up into preassembled
capsids. We have recently provided the first evidence that HSV
capsids contain disulfide bonds which may be important for viral
assembly and encapsidation. This rotation project will involve
the introduction of mutations into the conserved cysteine
residues of capsid proteins to determine whether proper
disulfide bond formation is important for during infection.
Selected Publications
Livingston, C. M., N. Deluca, D. E. Wilkinson and S. K. Weller.
2008. The formation of foci of ICP8, the single strand DNA
binding protein of HSV-1, requires the oligomerization of ICP4.
J. of Virol. 82:6324-36.
Chattopadhyay, S. and S.K. Weller. 2007. Direct interaction
between the N- and C-terminal portions of the Herpes Simplex
Virus Type 1 origin binding protein UL9 implies the formation of
a head-to-tail dimer. J Virol. 81:13659-67.
Iwahori S, N. Shirata, Y., Kawaguchi , S.K. Weller, Y. Sato, A.
Kudoh, S. Nakayama, H. Isomura, and T. Tsurumi. 2007. Enhanced
phosphorylation of transcription factor sp1 in response to
herpes simplex virus type 1 infection is dependent on the ataxia
telangiectasia-mutated protein. J Virol. 81:9653-64.
Nellissery, J.K, Szczepaniak, R., Lamberti, C. and S.K. Weller
2007. A putative leucine zipper within the HSV-1 UL6 protein is
required for portal ring formation. J. of Virol. 81:8868-77.
Chen, Y., C. M. Livingson, S. D. Carrington-Lawrence, P. Bai,
and S. K. Weller. 2007. A Mutation in the Human Herpes Simplex
Virus Type I UL52 Zinc Finger Motif Results in Defective Primase
Activity but Can Recruit Viral Polymerase and Support Viral
Replication Efficiently. J. of Virol. 81:8742-51
Chen, Y. Bai, P., Korza, G. Carson J.H. And S.K. Weller.
2009. The HSV-1 helicase-primase requires ssDNA to load and
unwind. Manuscript in revision.
Saffran, H.A., Pare, J.M. Corcoran, J.A., Weller, S.K. and
J.R. Smiley. 2007. Herpes simplex virus eliminates host
mitochondrial DNA. Embo J. 8:188-93.
Wilkinson, D. E., and S. K. Weller. 2006. Herpes Simplex
Virus Type I Disrupts the ATR-Dependent DNA Damage Response
During Lytic Infection. Journal of Cell Science J Cell Sci.
119:2695-703.
Chattopadhay, S., and Chen, Y., SK Weller. 2006. The two
helicases of herpes simplex virus type 1 (HSV-1). Front Biosci.
2006;11:2213-23.
Chattopadhay, S., and SK Weller. 2006. The DNA binding
activity of the HSV-1 origin binding protein, UL9, can be
modulated by sequences in the N-terminus: Correlation between
transdominance and DNA binding. J. Virol. 80:4491-500.
Burch A.D. and S.K. Weller. 2005 The HSV-1 DNA polymerase
requires the mammalian chaperone HSP90 for proper localization
to the nucleus. J. of Virol. 79:10740-9.
Pierce, AT, J. DeSalvo, TP Foster, A. Kosinski A, SK WelIer,
and WP Halford. 2005. nterferon-b and interferon-g synergize to
repress herpes simplex virus type 1 DNA synthesis and
nucleocapsid assembly. J. of Virol., 2005. J Gen Virol ; 86:
2421-2432.
Chen, Y. S.D. Carrington-Lawrence, P. Bai and S.K. Weller.
2005. Mutations in the Putative Zinc-Binding Motif of UL52
Demonstrate a Complex Interdependence between the UL5 and UL52
Subunits of the Human Herpes Simplex Virus Type 1 Helicase/Primase
Complex. J. Virol. 29:9088-9096.
Reuven, N.B. and S.K. Weller 2005 The Herpes simplex virus
type 1 single strand DNA binding protein ICP8 enhances the
nuclease activity of the UL12 alkaline nuclease by increasing
its processivity. J. Virol. 79: 9356-9358.
Wilkinson, D. E., and S. K. Weller. 2005. Inhibition of the
HSV-1 DNA Polymerase Induces the Hyperphosphorylation of
Replication Protein A and Its Accumulation at S Phase-specific
Sites of DNA Damage During Infection. J. Virol. 79: 7162-71.
Reuven, N.B., S. Willcox, J.D. Griffith, and S.K. Weller,
2004 Catalysis of strand exchange by the HSV-1 UL12 and ICP8
proteins: potent ICP8 recombinase activity is revealed upon
resection of dsDNA substrate by nuclease. Journal of Molecular
Biology, 342:57-71.
Burch A. and Weller SK. 2004. Nuclear sequestration of
cellular chaperone and proteasomal machinery during HSV-1
infection. J Virol,. 78(13): p. 7175-85.
Reuven, NB, Antoku, S, and Weller SK. 2004. UL12.5 retains
all known enzymatic functions of UL12 including strand exchange
but is unable to substitute for the lack of UL12 because it
lacks a nuclear localization signal. J Virol, 2004. 78(9): p.
4599-608.
Wilkinson, D. E., and S. K. Weller. 2004. Recruitment of
Cellular Recombination and Repair Proteins to Sites of HSV-1 DNA
Replication is Dependent upon the Composition of Viral Proteins
within Prereplicative Sites and Correlates with the Induction of
the DNA Damage Response. J. of Virol. 78:4783-96.
Wilkinson, D. E., and S. K. Weller. 2003. The Role of DNA
Recombination in Herpes Simplex Virus DNA Replication. IUBMB
Life 2003 55: 451-458.
Marintcheva, B. and S. K. Weller, 2003. Transdominant and
Potentiating Mutants of UL9, the HSV-1 origin binding protein,
suggest that levels of UL9 protein may be regulated during
infection. J. Virol. 77: 9630-51.
Prezch, A.J., D. Yu and Weller, S.K. 2003. Point mutations in
exon I of the Herpes Simplex Virus putative terminase subunit,
UL15, indicate that the most conserved residues are essential
for cleavage and packaging. J. of Virol. 77: 9613-21.
Reuven, N.B., Staire, A.E., Myers, R.S., and Weller, S.K.
2003. The Herpes Simplex Virus-1 Alkaline Nuclease and
Single-strand DNA Binding Protein Mediate Strand Exchange in
vitro. J Virol 77: 7425-33.
Carrington-Lawrence, S. D., and Weller, S. K. 2003.
Recruitment of Polymerase to Herpes Simplex Virus Type 1
Replication Foci in Cells Expressing Mutant Primase (UL52)
Proteins. J Virol 77: 4237-4247.
Marintcheva, B. and S. K. Weller. 2003. Helicase motif Ia is
involved in single-strand DNA-binding and helicase activities of
the herpes simplex virus type 1 origin-binding protein, UL9. J
Virol 77: 2477-88 |
