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Sequence elements essential for rho-dependent transcription termination at lambda tR1.

C Y Chen, J P Richardson

The Journal of Biological Chemistry

|August 15, 1987

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Researchers identified specific DNA sequences, rutA and rutB, crucial for rho factor-dependent transcription termination at the phage lambda tR1 terminator. These sequences are essential for rho factor binding and termination, unlike other contributing sequences.

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

Bacterial transcription termination is a complex process involving various factors and DNA sequences.
The rho factor is a key protein involved in transcription termination in many bacterial systems.
Understanding the precise DNA sequences that mediate rho factor action is crucial for deciphering gene regulation.

Purpose of the Study:

To pinpoint the specific DNA sequences essential for rho factor utilization at the phage lambda tR1 terminator.
To elucidate the role of these sequences in the interaction between rho factor and nascent RNA during transcription termination.

Main Methods:

Construction and analysis of deletion mutants within the cro-gene sequence of phage lambda.
In vitro transcription assays to assess the impact of mutations on rho factor-dependent termination.

Related Experiment Videos

Biochemical analysis of RNA-DNA interactions and inhibition studies using DNA oligonucleotides.

Main Results:

Two distinct sequence blocks, rutA and rutB, were identified as critical for rho factor action at the tR1 terminator.
These rut sequences are located between the cro mRNA stop codon and the tR1 terminator.
The rut sequences encode RNA segments rich in cytidylate residues and lacking extensive base pairing, suggesting a direct role in rho factor binding.
NusA protein's effect on rho action was examined, with deletions in the boxA sequence not affecting NusA's ability to reduce rho action.

Conclusions:

The rutA and rutB sequences are essential components of the recognition site for rho factor at the phage lambda tR1 terminator.
The identified rut transcript segments likely serve as the binding site for rho factor on the nascent cro RNA.
These findings provide critical insights into the mechanism of rho-dependent transcription termination.