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Journal of Bacteriology, January 2000, p. 456-462, Vol. 182, No. 2
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

Rodney A. King,* Peter L. Madsen,dagger and Robert A. Weisberg

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 lambda . Lambda resistance is caused by the action of the HK022 Nun protein, which prematurely terminates early lambda  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.

dagger Present address: Carlsberg Laboratory, Department of Yeast Genetics, 2500 Valby, Denmark.


Journal of Bacteriology, January 2000, p. 456-462, Vol. 182, No. 2
0021-9193/00/$04.00+0



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