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Published on: October 14, 2011
DNA methyltransferase-dependent transcription of the phage Mu mom gene
Abstract:
The phage Mu mom gene controls an unusual DNA modification. Expression of the mom function requires an active host (dam+) DNA adenine methylase [S-adenosyl-L-methionine:DNA (6-aminopurine)-methyltransferase]; in dam- hosts, Mu development is normal except that the viral DNA does not undergo the mom modification. The present communication compares transcription of the mom gene in dam+ versus dam- cells. 32P-labeled probes were prepared by nick-translation of a purified mom gene-containing restriction fragment and of virion DNA, respectively. These probes were hybridized with various RNAs blotted onto nitrocellulose filters (after fractionation by agarose gel electrophoresis). The salient findings are: (i) mom-specific RNA was readily detected in dam+ lysogenic cells, but only after induction of the Mu prophage; (ii) the level of mom RNA was decreased at least to 1/20th in induced dam- Mu lysogens; and (iii) little difference, if any, was observed between dam+ and dam- cells with respect to total Mu transcripts produced after prophage induction. These results are in accord with the known pattern of mom gene expression and Mu development. They show that the host (dam+) DNA adenine methylase activity is required for transcription of the mom gene. This represents a unique example where a DNA methylase exerts a positive regulatory role in mRNA transcription; alternative mechanisms for this process will be discussed.
Insights
Host DNA adenine methylase activity is crucial for phage Mu mom gene transcription. This DNA modification enzyme positively regulates mom gene mRNA levels, a unique regulatory role in gene expression.
Area of Science:
- Bacteriophage genetics
- Molecular biology
- Epigenetics
Background:
- Phage Mu mom gene controls a unique DNA modification.
- This modification requires host DNA adenine methylase (dam+).
- In dam- hosts, Mu development is normal but lacks mom modification.
Purpose of the Study:
- Compare mom gene transcription in dam+ versus dam- cells.
- Investigate the role of DNA adenine methylase in mom gene expression.
- Determine if dam+ activity is required for mom transcription.
Main Methods:
- Used 32P-labeled probes for mom gene and virion DNA.
- Hybridized probes with RNA blotted onto nitrocellulose filters.
- Fractionated RNA using agarose gel electrophoresis.
Main Results:
- Mom-specific RNA detected in induced dam+ lysogens.
- Mom RNA levels decreased at least 20-fold in induced dam- lysogens.
- Total Mu transcripts showed little difference between dam+ and dam- cells.
Conclusions:
- Host DNA adenine methylase activity is required for mom gene transcription.
- This demonstrates a unique positive regulatory role for DNA methylase in mRNA transcription.
- Alternative mechanisms for this regulatory process are discussed.
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