# Structural and mechanistic insights into AAV mediated site-specific integration

> **NIH NIH R01** · VIRGINIA COMMONWEALTH UNIVERSITY · 2020 · $298,625

## Abstract

PROJECT SUMMARY
Adeno-Associated Virus (AAV) Rep proteins use a unique mechanism to integrate into its host genome using a
strand-transfer reaction and also to process single stranded DNA during DNA replication. The large AAV Rep
proteins Rep68/Rep78 are the key players that carry out this process using two multifunctional domains that
include an origin binding domain (OBD) that binds DNA specifically and belongs to the family of HUH
endonucleases; and a SF3 helicase domain. Integration is contingent on the binding of Rep68/Rep78 to the
integration site AAVS1, promote its melting and perform a strand-specific transesterification reaction in the
single-stranded region that has been extruded during the melting step. Little is known about the molecular
details of this process and our long-term goal is to determine the molecular mechanism of site-specific
integration mediated by AAV Rep proteins. In this proposal we will focus on uncovering the mechanisms of
DNA melting and nicking using a combination of structural biology methods that include X-ray crystallography,
small-angle X-ray scattering (SAXS) and single-particle reconstruction electron microscopy coupled to
biochemical, biophysical and in vivo assays.

## Key facts

- **NIH application ID:** 9985906
- **Project number:** 5R01GM124204-04
- **Recipient organization:** VIRGINIA COMMONWEALTH UNIVERSITY
- **Principal Investigator:** Carlos R Escalante
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $298,625
- **Award type:** 5
- **Project period:** 2017-09-30 → 2021-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9985906

## Citation

> US National Institutes of Health, RePORTER application 9985906, Structural and mechanistic insights into AAV mediated site-specific integration (5R01GM124204-04). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/9985906. Licensed CC0.

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