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Journal of Bacteriology, January 2000, p. 456-462, Vol. 182, No. 2
Laboratory of Molecular Genetics, National
Institute of Child Health and Human Development, National Institutes of
Health, Bethesda, Maryland
Received 6 August 1999/Accepted 26 October 1999
Lysogens of phage HK022 are resistant to infection by phage
0021-9193/00/$04.00+0
Constitutive Expression of a Transcription
Termination Factor by a Repressed Prophage: Promoters for Transcribing
the Phage HK022 nun Gene
and
.
Lambda resistance is caused by the action of the HK022 Nun protein,
which prematurely terminates early
transcripts. We report here that
transcription of the nun gene initiates at a constitutive
prophage promoter, PNun, located just upstream
of the protein coding sequence. The 5' end of the transcript was determined by primer extension analysis of RNA isolated from HK022 lysogens or RNA made in vitro by transcribing a template containing the
promoter with purified Escherichia coli RNA polymerase.
Inactivation of PNun by mutation greatly
reduced Nun activity and Nun antigen in an HK022 lysogen. However, a
low level of residual activity was detected, suggesting that a
secondary promoter also contributes to nun expression. We
found one possible secondary promoter, PNun', just upstream of PNun. Neither promoter is
likely to increase the expression of other phage genes in a lysogen
because their transcripts should be terminated downstream of
nun. We estimate that HK022 lysogens in stationary phase
contain several hundred molecules of Nun per cell and that cells in
exponential phase probably contain fewer.
*
Corresponding author. Mailing address: Bldg. 6B, Room
408, 6 Center Dr., MSC 2785, NIH, Bethesda, MD 20892-2785. Phone: (301) 496-5663. Fax: (301) 496-0243. E-mail:
rking{at}lmgvax.nichd.nih.gov.
Present address: Carlsberg Laboratory, Department of Yeast
Genetics, 2500 Valby, Denmark.
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