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Acinetobacter baumannii encodes multiple mutagenically and biochemically active DNA polymerase V variants.

Debika Ojha1, Malgorzata Jaszczur1, Phuong Pham1

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Acinetobacter baumannii possesses multiple DNA polymerase V variants that cause mutations in E. coli. Mutagenesis requires RecA and specific polymerase components, highlighting complex DNA repair mechanisms.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Acinetobacter baumannii harbors multiple umuDC homologs, encoding diverse DNA polymerase V (pol V) variants.
  • These homologs suggest complex roles in DNA repair and mutagenesis within A. baumannii.

Purpose of the Study:

  • To investigate the mutagenic potential of A. baumannii pol V variants in E. coli.
  • To characterize the assembly and activity of A. baumannii pol V mutasomes.

Main Methods:

  • Expression of A. baumannii umuDC homologs in E. coli.
  • In vitro assembly and DNA synthesis assays of mutasome complexes.
  • Analysis of translesion DNA synthesis (TLS) at thymine-thymine cyclobutane dimers.

Main Results:

  • Six of twelve A. baumannii pol V variants exhibited mutagenic activity in E. coli, dependent on RecA.
  • Five active pol VAb variants formed stable mutasome complexes (UmuD'2CAb-RecAAb-ATP/ATPγS).
  • One mutasome required an A. baumannii-encoded β/τ clamp; TLS showed variant-specific misincorporation at T^T dimers.

Conclusions:

  • A. baumannii pol V variants contribute to mutagenesis, with specific variants and RecA being essential.
  • Mutasome complex formation and accessory factors like the β/τ clamp influence polymerase activity.
  • RecA's conserved role in mutagenesis is further supported by a specific mutation abolishing A. baumannii pol V activity.