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Eukaryotic Cell, June 2004, p. 579-588, Vol. 3, No. 3
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.3.579-588.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Novel Promoter Sequence Required for Manganese Regulation of Manganese Peroxidase Isozyme 1 Gene Expression in Phanerochaete chrysosporium

Biao Ma, Mary B. Mayfield, Bruce J. Godfrey, and Michael H. Gold^*

Department of Biochemistry and Molecular Biology, OGI School of Science and Engineering, Oregon Health & Science University, Beaverton, Oregon 97006-8921

Received 8 December 2003/ Accepted 31 March 2004

Manganese peroxidase (MnP) is a major, extracellular component of the lignin-degrading system produced by the wood-rotting basidiomycetous fungus Phanerochaete chrysosporium. The transcription of MnP-encoding genes (mnps) in P. chrysosporium occurs as a secondary metabolic event, triggered by nutrient-nitrogen limitation. In addition, mnp expression occurs only under Mn²⁺ supplementation. Using a reporter system based on the enhanced green fluorescent protein gene (egfp), we have characterized the P. chrysosporium mnp1 promoter by examining the effects of deletion, replacement, and translocation mutations on mnp1 promoter-directed egfp expression. The 1,528-bp mnp1 promoter fragment drives egfp expression only under Mn²⁺-sufficient, nitrogen-limiting conditions, as required for endogenous MnP production. However, deletion of a 48-bp fragment, residing 521 bp upstream of the translation start codon in the mnp1 promoter, or replacement of this fragment with an unrelated sequence resulted in egfp expression under nitrogen limitation, both in the absence and presence of exogenous Mn²⁺. Translocation of the 48-bp fragment to a site 120 bp downstream of its original location resulted in Mn²⁺-dependent egfp expression under conditions similar to those observed with the wild-type mnp1 promoter. These results suggest that the 48-bp fragment contains at least one Mn²⁺-responsive cis element. Additional promoter-deletion experiments suggested that the Mn²⁺ element(s) is located within the 33-bp sequence at the 3' end of the 48-bp fragment. This is the first promoter sequence containing a Mn²⁺-responsive element(s) to be characterized in any eukaryotic organism.

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* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, OGI School of Science and Engineering, Oregon Health & Science University, 20000 N.W. Walker Rd., Beaverton, OR 97006-8921. Phone: (503) 646-0957. Fax: (503) 748-1464. E-mail: mhgold{at}myexcel.com.

Present address: Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205.

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Eukaryotic Cell, June 2004, p. 579-588, Vol. 3, No. 3
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.3.579-588.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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