# VSV STRUCTURE, BUDDING, ENTRY USING MEMBRANE MUTANTS

> **NIH NIH R01** · UNIV OF MED/DENT NJ-R W JOHNSON MED SCH · 1988 · $237,143

## Abstract

The aim of this research is to describe as completely as possible the
action of M protein during that part of the infectious cycle commencing
with virion budding from an infected cell, and concluding with the release
of the genome-containing necleocapsid in a transcriptionally active form in
the cytoplasma of an uninfected cell.  Previously characterized temperature
sensitive M protein mutants of VSV will be used in this study, and also VS
virions made transcriptionally active by addition of melittin.  The
sequence alterations that confer altered function on the M protein mutants
will be determined by sequence analysis of cDNA prepared using the mutant
genome as template.  The identity of the alteration(s) will be confirmed by
sequencing revertants of these mutants.  The infectivity of intact and
melittin permeabilized VSV will be compared, using both mutant and wild
type VSV.  The proteins of melittin permeabilized VSV will be
phosphorylated to determine whether this affects infectvity.  The effect of
phosphorylation of M on its nucleocapsid-binding and membrane-binding
properties will be assessed, using melittin permeabilized VSV.  M in
different phosphorylated forms will be added to functioning nucleocapsids
in detergent to assess the effect of phosphorylation on inhibition of
transcription by M.  The effect of M protein phosphorylation of G protein
mobility following fusion will be assessed using fluorescence
photobleaching recovery (FPR) on erythrocytes fused with VSV.  FPR
experiments will also be carried out using giant liposomes as target
membranes; permeabilization of these with melittin might permit the study
of G-M protein interactions in a completely reconstituted system.  The
thermolability of the highly thermolabile M mutant ts G31 will be
investigated using melittin permeabilized virions to determine whether
thermal inactivation blocks phosphorylation of M, thus blocking onset of
transcription.  If phosphorylation does alter the functional properties of
M, phosphorylation sites will be identified by peptide analysis, and the
protein kinase will be purified and characterized from virions.  Attempts
will be made to prepare fragments of M by limited proteolytic digestion;
these will be assessed for nuecleocapsid and membrane binding activities,
and as substrates for phosphorylation.

## Key facts

- **NIH application ID:** 3125351
- **Project number:** 5R01AI013002-13
- **Recipient organization:** UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
- **Principal Investigator:** JOHN LENARD
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $237,143
- **Award type:** 5
- **Project period:** 1976-09-01 → 1989-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3125351

## Citation

> US National Institutes of Health, RePORTER application 3125351, VSV STRUCTURE, BUDDING, ENTRY USING MEMBRANE MUTANTS (5R01AI013002-13). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3125351. Licensed CC0.

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