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Functionally distinct, sequence-specific replicator and origin elements are required for Drosophila chorion gene

L Lu1, H Zhang, J Tower

  • 1Department of Biological Sciences, University of Southern California, Los Angeles, California 90089-1340, USA.

Genes & Development
|February 7, 2001
PubMed
Summary
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Drosophila follicle cells amplify chorion genes using an amplification control element (ACE3) that activates a DNA replication origin (ori-beta) in cis, supporting the replicator model for gene amplification.

Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Genetics

Background:

  • Drosophila ovarian follicle cells amplify chorion gene clusters up to 60-fold to meet protein synthesis demands.
  • Amplification relies on repeated firing of DNA replication origins within gene clusters.

Purpose of the Study:

  • To investigate the sequence requirements for chorion gene amplification.
  • To identify the functional roles of the amplification control element (ACE3) and ori-beta in DNA replication initiation.

Main Methods:

  • Deletion analyses of transgenic constructs.
  • Two-dimensional (2D) gel electrophoresis to identify DNA replication origins.
  • Use of a buffered vector with insulator elements (SHWBS) to mitigate chromosomal position effects.

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Main Results:

  • The 320-bp ACE3 and 884-bp ori-beta elements were found to be necessary and sufficient for amplification.
  • ori-beta functioned as the DNA replication origin, while ACE3 did not show origin activity.
  • An insulator between ACE3 and ori-beta inhibited amplification, indicating ACE3 activates ori-beta in cis.

Conclusions:

  • ACE3 acts as a replicator, activating ori-beta to initiate DNA replication.
  • These findings support and extend the replicator model for metazoan chromosomal replicons.