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Journal of Bacteriology, April 2003, p. 2338-2345, Vol. 185, No. 7
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.7.2338-2345.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Streptomyces coelicolor Developmental Transcription Factor ^BldN Is Synthesized as a Proprotein

Maureen J. Bibb and Mark J. Buttner^*

Department of Molecular Microbiology, John Innes Centre, Colney, Norwich NR4 7UH, United Kingdom

Received 25 October 2002/ Accepted 9 January 2003

bldN is one of a set of genes required for the formation of specialized, spore-bearing aerial hyphae during differentiation in the mycelial bacterium Streptomyces coelicolor. Previous analysis (M. J. Bibb et al., J. Bacteriol. 182:4606-4616, 2000) showed that bldN encodes a member of the extracytoplasmic function subfamily of RNA polymerase factors and that translation from the most strongly predicted start codon (GTG¹) would give rise to a factor having an unusual N-terminal extension of ca. 86 residues. Here, by using a combination of site-directed mutagenesis and immunoblot analysis, we provide evidence that all bldN translation arises from initiation at GTG¹ and that the primary translation product is a proprotein (pro-^BldN) that is proteolytically processed to a mature species (^BldN) by removal of most of the unusual N-terminal extension. A time course taken during differentiation of the wild type on solid medium showed early production of pro-^BldN and the subsequent appearance of mature ^BldN, which was concomitant with aerial mycelium formation and the disappearance of pro-^BldN. Two genes encoding members of a family of metalloproteases that are involved in the regulated proteolytic processing of transcription factors in other organisms were identified in the S. coelicolor genome, but their disruption did not affect differentiation or pro-^BldN processing.

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* Corresponding author. Mailing address: Department of Molecular Microbiology, John Innes Centre, Colney, Norwich NR4 7UH, United Kingdom. Phone: (44)(0)1603-450759. Fax: (44)(0)1603-450778. E-mail: mark.buttner{at}bbsrc.ac.uk.

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Journal of Bacteriology, April 2003, p. 2338-2345, Vol. 185, No. 7
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.7.2338-2345.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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