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Transcription termination and processing sites in the bacteriophage lambda pL operon.

H C Hyman, A Honigman

    Journal of Molecular Biology
    |May 5, 1986
    PubMed
    Summary

    Bacteriophage lambda

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    Area of Science:

    • Molecular Biology
    • Genetics
    • Virology

    Background:

    • The bacteriophage lambda major leftward operon (PL operon) is crucial for viral gene expression.
    • Understanding transcriptional termination and processing within this operon is key to viral replication.
    • Previous studies identified some termination sites but lacked comprehensive mapping of all stable RNA 3' ends.

    Purpose of the Study:

    • To precisely map the 3' ends of stable RNA species within the bacteriophage lambda PL operon.
    • To investigate the roles of the N antitermination function, Rho factor, and RNase III in transcript termination and processing.
    • To elucidate the mechanisms controlling gene expression within the PL operon.

    Main Methods:

    • S1 nuclease mapping was employed to analyze transcripts.
    • RNA was purified from induced lambda prophage and bacterial plasmids containing the PL operon.
    • Experiments were conducted under various conditions manipulating N function, Rho factor, and RNase III activity.

    Main Results:

    • Three transcript termination sites were identified in the N-Rho+ intercistronic region (positions 34,826, 34,558, and 34,393).
    • Additional RNA species with 3' ends mapped between the ral and Ea10 genes, with one site showing partial N-resistance.
    • RNase III deficiency and N function influenced transcript processing, creating distinct 3' ends at positions 33,910 and 33,980.

    Conclusions:

    • Transcriptional elongation in the lambda PL operon is regulated by sequential termination events between genes.
    • Post-transcriptional processing, particularly by RNase III and influenced by the N protein, plays a significant role in controlling gene expression.
    • These findings reveal a complex regulatory network governing lambda PL operon expression.

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