Genetic dissection of T4 lysis.

Abstract:

:t is the holin gene for coliphage T4, encoding a 218-amino-acid (aa) protein essential for the inner membrane hole formation that initiates lysis and terminates the phage infection cycle. T is predicted to be an integral membrane protein that adopts an N(in)-C(out) topology with a single transmembrane domain (TMD). This holin topology is different from those of the well-studied holins S105 (3 TMDs; N(out)-C(in)) of the coliphage lambda and S68 (2 TMDs; N(in)-C(in)) of the lambdoid phage 21. Here, we used random mutagenesis to construct a library of lysis-defective alleles of t to discern residues and domains important for holin function and for the inhibition of lysis by the T4 antiholin, RI. The results show that mutations in all 3 topological domains (N-terminal cytoplasmic, TMD, and C-terminal periplasmic) can abrogate holin function. Additionally, several lysis-defective alleles in the C-terminal domain are no longer competent in binding RI. Taken together, these results shed light on the roles of the previously uncharacterized N-terminal and C-terminal domains in lysis and its real-time regulation.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Moussa SH,Lawler JL,Young R

doi

10.1128/JB.01548-14

subject

Has Abstract

pub_date

2014-06-01 00:00:00

pages

2201-9

issue

12

eissn

0021-9193

issn

1098-5530

pii

JB.01548-14

journal_volume

196

pub_type

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