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TraM of plasmid R1 regulates its own expression | JoVE Visualize

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TraM of plasmid R1 regulates its own expression

M Schwab¹, H Reisenzein, G Högenauer

¹Institut für Mikrobiologie, Universität Graz, Austria.

Molecular Microbiology|March 1, 1993

View abstract on PubMed

Summary

The TraM protein autoregulates its own gene expression (traM) in resistance plasmid R1. This regulation requires the N-terminus of TraM and is allele-specific, impacting DNA binding.

Area of Science:

Molecular Biology
Microbiology
Genetics

Background:

The traM gene is crucial for the conjugation of resistance plasmid R1.
Understanding gene regulation is key to controlling plasmid transfer and antibiotic resistance dissemination.

Purpose of the Study:

To investigate the in vivo regulation of the traM gene.
To determine the role of the TraM protein in its own gene's expression.
To identify the specific protein domains involved in TraM-mediated regulation.

Main Methods:

Utilized translational gene fusion (traM-lacZ) to monitor traM expression.
Integrated fusion constructs into the bacterial chromosome using recombinant phage lambda RZ5.
Employed site-specific mutagenesis to probe the function of the TraM N-terminus.

Main Results:

The traM gene exhibits highly active promoters.
Presence of R1 plasmids in trans significantly reduced traM expression (5-45 fold), indicating autoregulation.
Mutagenesis of the TraM N-terminus abolished autoregulation, highlighting its importance in DNA binding.
Autoregulation was found to be allele-specific, with TraM recognizing cognate DNA.

Conclusions:

The TraM protein autoregulates traM gene expression.
The N-terminal region of TraM is essential for its DNA-binding function in autoregulation.
TraM autoregulation is allele-specific, suggesting distinct DNA recognition mechanisms.

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Tram Of Plasmid R1 Regulates Its Own Expression