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DNA helical instability facilitates initiation at the SV40 replication origin

S Lin1, D Kowalski

  • 1Molecular and Cellular Biology Department, Roswell Park Cancer Institute, Buffalo, NY 14263.

Journal of Molecular Biology
|January 14, 1994
PubMed
Summary

The SV40 replication origin

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA replication origins require specific elements for function.
  • A DNA unwinding element (DUE) with intrinsic helical instability is crucial for bacterial and yeast origins.

Purpose of the Study:

  • To investigate the role of helical instability in the Simian virus 40 (SV40) replication origin.
  • To determine if helical instability contributes to SV40 origin function in vitro and in vivo.

Main Methods:

  • Analyzing mutations in SV40 replication origin sequences.
  • Assessing the helical stability of mutant origin sequences.
  • Measuring origin activity in vitro and in vivo.

Main Results:

  • Both the early palindrome (EP) and A+T-rich (AT) domains of the SV40 origin exhibit helical instability.
  • Origin activity inversely correlates with helical stability of mutations in the EP domain, but not the AT or T-antigen binding domains.
  • A specific EP subdomain's helical instability is essential for T-antigen-induced DNA melting and replication initiation.

Conclusions:

  • Helical instability in a specific EP subdomain acts as the DUE for the SV40 replication origin.
  • This instability is critical for facilitating T-antigen binding and subsequent DNA strand separation.
  • The findings define the DUE of the SV40 origin based on its sensitivity to helix-stabilizing mutations.

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