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Functional analysis of the phage T4 holin in a lambda context.

E Ramanculov¹, R Young

¹Department of Biochemistry and Biophysics, Texas A and M University, College Station 77843-2128, USA.

Molecular Genetics and Genomics : MGG

|May 22, 2001

Summary

Phage T4's t gene functions as a holin, controlling lysis timing. Combining T4 and lambda holins accelerates lysis, suggesting a shared pathway for holin function.

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Area of Science:

Bacteriophage biology
Molecular genetics
Cell lysis mechanisms

Background:

Bacteriophages employ holin proteins to trigger host cell lysis.
The precise mechanisms and evolutionary conservation of holin function remain under investigation.
Phage lambda and T4 possess distinct holin genes (S and t, respectively).

Purpose of the Study:

To functionally characterize the T4 phage t gene as a holin.
To investigate the regulation of lysis inhibition (LIN) by T4 holin.
To explore the combined effects of lambda and T4 holins on lysis timing.

Main Methods:

Construction of lambda-T4 hybrid phages with T4 t gene replacing lambda S gene.
Induction of hybrid and parental phages to observe lysis timing.
Superinfection experiments with LIN-competent T4 phage to study lysis inhibition.
Analysis of holin gene allelic states in superinfecting T4 phage.

Main Results:

The T4 t gene functions as a holin, inducing abrupt lysis earlier than wild-type lambda.
Lysis inhibition (LIN) could be imposed on hybrid phage induction by T4 superinfection.
LIN imposition depended on T4 rI and t gene alleles, indicating a transient sensitive state for T4 holin.
Co-induction of lambda and T4 holins resulted in earlier lysis compared to individual holins.

Conclusions:

The T4 t gene is a functional holin gene.
T4 holin exhibits a transient sensitivity to lysis inhibition.
Lambda and T4 holins likely act through a common pathway, potentially additively influencing lysis timing.

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