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Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters

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Published on: November 26, 2013

DNA binding is rate-limiting for natural transformation.

Taylor J Ellison¹, Courtney K Ellison¹

¹Department of Microbiology, University of Georgia, Athens, GA.

Biorxiv : the Preprint Server for Biology

|June 19, 2024

Summary

Bacteria use type IV pili (T4P) for natural transformation, a key horizontal gene transfer process. Enhanced DNA binding by the FimT pilin in Acinetobacter baylyi significantly boosts this bacterial gene transfer capability.

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Published on: August 21, 2018

Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Natural transformation is a widespread bacterial horizontal gene transfer mechanism.
Factors limiting natural transformation across bacterial species remain poorly understood.
Type IV pili (T4P) are essential for DNA uptake during natural transformation.

Purpose of the Study:

To investigate the molecular basis for the high natural transformability of *Acinetobacter baylyi*.
To identify factors contributing to or limiting DNA binding by T4P.
To assess the role of T4P-DNA binding efficiency in natural transformation.

Main Methods:

Comparative analysis of FimT pilin sequences and structures.
Biochemical assays to measure DNA binding efficiency.
Genetic manipulation of *Acinetobacter* species to express heterologous FimT.

Main Results:

*Acinetobacter baylyi* FimT exhibits enhanced nonspecific DNA binding due to multiple positively charged residues.
This enhanced DNA binding by FimT significantly increases natural transformation efficiency.
Expression of *A. baylyi* FimT in a related pathogen improves its transformation capacity.

Conclusions:

T4P-DNA binding efficiency is a critical determinant of natural transformation.
FimT's unique properties in *A. baylyi* explain its high transformability.
Targeting T4P-DNA interactions could modulate horizontal gene transfer in bacteria.