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Related Experiment Videos

The role of the OOP antisense RNA in coliphage lambda development.

L Krinke1, M Mahoney, D L Wulff

  • 1Department of Biological Sciences, State University of New York, Albany 12222.

Molecular Microbiology
|May 1, 1991
PubMed
Summary

Researchers created a mutant bacteriophage lambda that does not produce OOP antisense RNA. This mutation impacts phage replication and gene expression, particularly affecting cll mRNA levels after UV induction.

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Area of Science:

  • Molecular Biology
  • Virology
  • Genetics

Background:

  • Bacteriophage lambda is a well-studied virus with complex regulatory mechanisms.
  • Antisense RNAs play crucial roles in gene regulation in various biological systems.
  • The OOP antisense RNA in bacteriophage lambda has been implicated in regulating gene expression.

Purpose of the Study:

  • To investigate the function of OOP antisense RNA in bacteriophage lambda replication.
  • To characterize a mutant bacteriophage lambda (lambda po-) lacking functional OOP antisense RNA.
  • To determine the impact of OOP antisense RNA on host and phage gene expression, particularly cll mRNA and O mRNA.

Main Methods:

  • Construction of a point mutant bacteriophage lambda (lambda po-) with an inactivated OOP promoter (po).

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  • Comparative analysis of wild-type lambda (+) and lambda po- phages regarding plaque morphology, growth curves, and lysogenization frequency.
  • Assessment of phage burst size following ultraviolet (UV) induction.
  • Analysis of cll mRNA and O mRNA cleavage events and levels during infection.

    • Main Results:

    • The lambda po- mutant exhibited identical plaque morphology, growth curves, and lysogenization frequencies compared to wild-type lambda (+).
    • UV induction resulted in a two-fold greater phage burst in lambda (+) infections than in lambda po- infections.
    • OOP RNA mediated two cleavage events in cll mRNA, leading to significantly lower cll mRNA levels in lambda (+) infections late in the cycle.
    • O mRNA levels were largely unaffected by the po- mutation.

    Conclusions:

    • The OOP antisense RNA is involved in regulating bacteriophage lambda replication, particularly evident after UV induction.
    • OOP RNA specifically targets and cleaves cll mRNA, impacting its stability and abundance.
    • The po- mutation provides a tool to study the specific role of OOP RNA in bacteriophage lambda gene regulation and infection dynamics.