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The chloroplast infA gene with a functional UUG initiation codon.

T Hirose1, T Ideue, T Wakasugi

  • 1Center for Gene Research, Nagoya University, Japan.

FEBS Letters
|March 9, 1999
PubMed
Summary

The green alga Chlorella vulgaris uses an unusual TTG start codon for its infA gene. While functional, this codon is inefficient for protein translation initiation.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Algal Biology

Background:

  • Chloroplast genes typically initiate translation using ATG start codons, with GTG as a rare alternative.
  • The infA gene in the green alga Chlorella vulgaris was identified as potentially using a non-canonical TTG start codon.

Purpose of the Study:

  • To investigate the function of the putative TTG initiation codon in the Chlorella vulgaris infA gene.
  • To determine the efficiency of translation initiation from this non-canonical codon.

Main Methods:

  • Reverse transcription-polymerase chain reaction (RT-PCR) was used to confirm transcription of the infA gene.
  • In vitro translation assays were performed using a tobacco chloroplast system to assess mRNA translation.
  • Site-directed mutagenesis was employed to change the TTG codon to ATG for comparison.

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Main Results:

  • Transcription of the infA gene in Chlorella vulgaris was confirmed.
  • The infA mRNA was accurately translated initiating from the TTG codon in a heterologous chloroplast system.
  • Mutating the TTG codon to ATG significantly increased translation efficiency by approximately 300-fold.

Conclusions:

  • The TTG codon is functional for accurate translation initiation of Chlorella vulgaris infA mRNA.
  • However, the TTG initiation codon is highly inefficient compared to the standard ATG codon.