Accession Number:

ADA458311

Title:

Maintaining Genome Stability: The Role of Helicases and Deaminases

Descriptive Note:

Annual rept. 1 Jul 2005-30 Jun 2006

Corporate Author:

UNIVERSITY OF SOUTHERN CALIFORNIA LOS ANGELES

Personal Author(s):

Report Date:

2006-07-01

Pagination or Media Count:

9.0

Abstract:

The Brief Description of The Four Aims For The Grant Aim 1. Study the in vitro functions of MCM proteins from archaea and yeast cells using the genetically engineered protein constructs. In this aim, we will also extend our prior success in the X-ray structural studies of an N-terminal fragment of an archaea MCM by attempting to crystallize MCM proteins from yeast. Aim 2. Examine in vivo effects of helicase function and in particular MCM roles in maintaining genome integrity in response to damage. This aim will use existing and newly generated mutants, which can be achieved through genetic screening and site-directed mutagenesis based on the 3-dimensional structure of MCM, to investigate how MCMs contribution to genome stability during chemical damage. Aim 3. Express, purify and crystallize the proteins of deaminases. We will focus on AID and APOBEC3G to obtain purified deaminase proteins for the in vitro biochemical, functional, and structural studies. Aim 4. Examine the functions and substrate specificity of AID and identify other factors required for the coupling of deamination with other processes of DNA synthesis and RNA transcription. The experiments will be carried out in a cell free assay system, using already purified DNA replicationrepair and RNA transcription proteins in our labs.

Subject Categories:

  • Biochemistry
  • Genetic Engineering and Molecular Biology
  • Medicine and Medical Research

Distribution Statement:

APPROVED FOR PUBLIC RELEASE