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Applied and Environmental Microbiology, February 1999, p. 569-577, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Identification and Characterization of a Lysis Module Present
in a Large Proportion of Bacteriophages Infecting
Streptococcus thermophilus
Michelle M.
Sheehan,1
Elizabeth
Stanley,1
Gerald F.
Fitzgerald,1,2,3 and
Douwe
van
Sinderen2,*
The National Food Biotechnology
Centre1 and
Departments of
Microbiology2 and
Food Science and
Technology,3 University College Cork, Cork,
Ireland
Received 27 April 1998/Accepted 29 October 1998
A lysis module encoded by the temperate bacteriophage
O1205 was
identified. This lysis module contains a lysin gene, designated lyt51, and two putative holin-encoding genes, designated
lyt49 and lyt50. lyt51 encodes a
lytic enzyme specifically directed against streptococcal cell walls.
Similar to other phage-encoded lysins, Lyt51 appears to have a modular
design in which the N-terminal portion corresponds to its enzymatic
activity while the C-terminal region is responsible for its substrate
binding specificity. The two putative holin-encoding genes,
lyt49 and lyt50, located immediately upstream
of lyt51, were identified on the basis of their homology to
other identified holin-encoding genes. Expression of lyt49 or lyt50 in Escherichia coli was shown to cause
cell death and leakage of the intracellular enzyme isocitrate
dehydrogenase into the growth medium without apparent lysis of the
cells. Southern blotting experiments demonstrated that at least one of
the three components of the identified lysis module is present in all
members of a large collection of bacteriophages, indicating that
components of this lysis module are widespread among bacteriophages
infecting Streptococcus thermophilus.
*
Corresponding author. Mailing address: Department of
Microbiology, University College Cork, Cork, Ireland. Phone: 353 21 902811. Fax: 353 21 903101. E-mail: douwe{at}ucc.ie.
Applied and Environmental Microbiology, February 1999, p. 569-577, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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