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Recurrent locus-specific mutation resulting from a cryptic ectopic insertion in Neurospora.

David D Perkins1, Michael Freitag, Virginia C Pollard

  • 1Department of Biological Sciences, Stanford University, Stanford, California 94305-5020, USA.

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|February 27, 2007
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Summary

The eas(UCLA) mutation induces new mutations in the cya-8 gene via repeat-induced point mutation (RIP). This occurs because an ectopic DNA insertion at eas acts as a locus-specific mutator.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Mycology

Background:

  • The eas allele UCLA191 (easily wettable, hydrophobin-deficient) is linked to genetic mutations.
  • Approximately 20% of progeny exhibit new mutations when parents carry the eas allele UCLA191.
  • These mutations inactivate the wild-type cya-8 allele, leading to thin, transparent mycelial growth.

Purpose of the Study:

  • To investigate the mechanism behind the recurrent cya-8 mutations observed in progeny of eas(UCLA) carriers.
  • To determine if repeat-induced point mutation (RIP) is responsible for the observed mutations.
  • To elucidate the role of ectopic DNA insertions in locus-specific mutagenesis.

Main Methods:

  • Crosses were performed between strains carrying the eas(UCLA) allele and wild-type strains.
  • Analysis of progeny for cya-8 mutations and their genetic linkage.
  • Investigation of the effects of unlinked duplications and RIP suppressors (rid) on mutation rates.
  • DNA analysis of the eas(UCLA) allele, including methylation patterns.

Main Results:

  • Recurrent cya-8 mutations are attributed to repeat-induced point mutation (RIP) caused by a duplicated cya-8+ copy ectopically inserted at the eas locus.
  • These RIP-induced mutations occur during nuclear proliferation before karyogamy.
  • Mutation frequency is influenced by genetic background, including unlinked duplications and RIP suppressors.
  • The eas(UCLA) allele contains a DNA discontinuity and its promoter is methylated in mutant progeny.

Conclusions:

  • The findings support a model where an ectopic DNA insertion at the eas locus acts as a locus-specific mutator through RIP.
  • Repeat-induced point mutation is a significant mechanism for generating mutations in this genetic context.
  • Understanding this mechanism provides insights into gene regulation and mutation processes.