DNA REPLICATION INITIATION SITES IN MAMMALIAN CELLS

Project: Research project

Project Details

Description

We will use two model systems to study DNA replication in human cell lines.
1. The role of the Epstein-Barr virus (EBV) nuclear antigen I (EBNA-1) in
the formation of a replication barrier at the EBV family of repeated
sequences in human cell lines will be studied using an in vitro replication
system. We will determine the minimum number of repeats that are required
to produce a barrier to replication in vitro and determine whether EBNA-1
binding to the family of repeats inhibits helicase activity of the T
antigen. We will then determine whether cellular proteins can also produce
a replication barrier by binding to the EBV family of repeats. 2. We will
identify replication origins in the rDNA gene cluster that replicate early
or late during the S phase.

We will determine the location of the sites at which the replication of the
immunoglobulin heavy chain multigene family initiates. This will be
determined in cell lines in which one or more of these genes are expressed
and in cell lines in which they are transcriptionally silent. We will use
deletion analysis to determine the sequences that are critical for these
immunoglobulin replication origins to function. Targeted integration
mediated by homologous recombination will be used to modify the chromosomal
origin region.
StatusFinished
Effective start/end date1/8/9212/31/92

Funding

  • National Institute of General Medical Sciences

ASJC

  • Genetics
  • Molecular Biology
  • Cancer Research
  • Virology
  • Cell Biology

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