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Journal of Bacteriology, May 2004, p. 3160-3172, Vol. 186, No. 10
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.10.3160-3172.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Organization and Expression of the Polynucleotide Phosphorylase Gene (pnp) of Streptomyces: Processing of pnp Transcripts in Streptomyces antibioticus

Patricia Bralley* and George H. Jones

Department of Biology, Emory University, Atlanta, Georgia 30322

Received 18 September 2003/ Accepted 29 January 2004

We have examined the expression of pnp encoding the 3'-5'-exoribonuclease, polynucleotide phosphorylase, in Streptomyces antibioticus. We show that the rpsO-pnp operon is transcribed from at least two promoters, the first producing a readthrough transcript that includes both pnp and the gene for ribosomal protein S15 (rpsO) and a second, Ppnp, located in the rpsO-pnp intergenic region. Unlike the situation in Escherichia coli, where observation of the readthrough transcript requires mutants lacking RNase III, we detect readthrough transcripts in wild-type S. antibioticus mycelia. The Ppnp transcriptional start point was mapped by primer extension and confirmed by RNA ligase-mediated reverse transcription-PCR, a technique which discriminates between 5' ends created by transcription initiation and those produced by posttranscriptional processing. Promoter probe analysis demonstrated the presence of a functional promoter in the intergenic region. The Ppnp sequence is similar to a group of promoters recognized by the extracytoplasmic function sigma factors, sigma-R and sigma-E. We note a number of other differences in rspO-pnp structure and function between S. antibioticus and E. coli. In E. coli, pnp autoregulation and cold shock adaptation are dependent upon RNase III cleavage of an rpsO-pnp intergenic hairpin. Computer modeling of the secondary structure of the S. antibioticus readthrough transcript predicts a stem-loop structure analogous to that in E. coli. However, our analysis suggests that while the readthrough transcript observed in S. antibioticus may be processed by an RNase III-like activity, transcripts originating from Ppnp are not. Furthermore, the S. antibioticus rpsO-pnp intergenic region contains two open reading frames. The larger of these, orfA, may be a pseudogene. The smaller open reading frame, orfX, also observed in Streptomyces coelicolor and Streptomyces avermitilis, may be translationally coupled to pnp and the gene downstream from pnp, a putative protease.

* Corresponding author. Mailing address: Department of Biology, 1510 Clifton Rd., Emory University, Atlanta, GA 30322. Phone: (404) 727-4208. Fax: (404) 727-2880. E-mail: pbralley{at}biology.emory.edu.

Journal of Bacteriology, May 2004, p. 3160-3172, Vol. 186, No. 10
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.10.3160-3172.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Chang, S. A., Cozad, M., Mackie, G. A., Jones, G. H. (2008). Kinetics of Polynucleotide Phosphorylase: Comparison of Enzymes from Streptomyces and Escherichia coli and Effects of Nucleoside Diphosphates. J. Bacteriol. 190: 98-106 [Abstract] [Full Text]  
  • Charaniya, S., Mehra, S., Lian, W., Jayapal, K. P., Karypis, G., Hu, W.-S. (2007). Transcriptome dynamics-based operon prediction and verification in Streptomyces coelicolor. Nucleic Acids Res 35: 7222-7236 [Abstract] [Full Text]  
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