# STRUCTURE AND FUNCTION OF VARICELLA ZOSTER DNA

> **NIH NIH R01** · U.S.UNIFORMED SERVICES UNIV  HLTH SCIS · 1989 · $66,847

## Abstract

This proposal aims to continue our studies of the molecular biology of
Varicella Zoster Virus (VZV).  We plan to pursue the investigation of the
structure of VZV DNA with special emphasis on the mode of formation and
frequency of occurrence of circular forms and forms with inverted UL
segments.  We will further characterize VZV DNA sequences from which strain
differences arise, particularly with reference to possible roles as
transcription signals.  We will attempt to map the VZV thymidine kinase
(TK) and DNA polymerase activities physically on the viral genome by rescue
of drug-resistant mutants and by use of special cell lines.  We will also
attempt to obtain expression of viral proteins from VZV sequences cloned
into mammalian virus expression systems including vaccinia and bovine
papilloma viruses.  Our initial target will be the 92K VZV glycoprotein.
We will purify and characterize the major 125K VZV DNA-binding protein,
comparing its DNA-binding and DNA polymerase-stimulating activities with
that of the HSV ICP8.  The genomic location of the 125K protein coding
sequences will be determined by marker rescue of HSV1 ICP8 ts mutants by
VZV fragments.  A similar strategy will be employed to attempt rescue of
HSV ts mutants in other important genes such as the TK, DNA polymerase and
major capsid proteins using the CaPO4 transfection scheme.  We also plan to
characterize further a virion-associated protein kinase activity in regard
to salt, detergent and substrate concentrations; the role of this kinase
activity in the formation of viral phosphoproteins and in virus infectivity
will be assessed.  Finally, we will continue our attempts to detect the
presence of VZV genetic material in human sensory ganglia.  We have
arranged for a new source of tissue and should be supplied with both
trigeminal and dorsal root ganglia.  The methods of detection will include
in situ hybridization using frozen thin sections of explanted ganglia, dot
blots and Southern blots.  Southern blots will also be used to examine the
conformation of detected sequences.  Techniques to be employed include
restriction enzyme analysis, CSC1 density gradient centrifugation, nick
translation, dot and Southern blotting, gel electrophoresis, DNA
transfection, filter binding, in situ hybridization and DNA sequencing.

## Key facts

- **NIH application ID:** 3127951
- **Project number:** 5R01AI018449-08
- **Recipient organization:** U.S.UNIFORMED SERVICES UNIV  HLTH SCIS
- **SAM.gov UEI:** Z83LJK5NWBC1
- **Principal Investigator:** WILLIAM T RUYECHAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1989
- **Award amount:** $66,847
- **Award type:** 5
- **Project period:** 1982-08-01 → 1990-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3127951

## Citation

> US National Institutes of Health, RePORTER application 3127951, STRUCTURE AND FUNCTION OF VARICELLA ZOSTER DNA (5R01AI018449-08). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3127951. Licensed CC0.

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