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Journal of Bacteriology, May 2006, p. 3208-3218, Vol. 188, No. 9
0021-9193/06/$08.00+0     doi:10.1128/JB.188.9.3208-3218.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

In Vitro and In Vivo Analysis of the Role of PrrA in Rhodobacter sphaeroides 2.4.1 hemA Gene Expression

Britton Ranson-Olson,1 Denise F. Jones,2 Timothy J. Donohue,2 and Jill H. Zeilstra-Ryalls1*

Department of Biological Sciences, Oakland University, Rochester, Michigan 48309,1 Department of Bacteriology, University of Wisconsin—Madison, Madison, Wisconsin 530762

Received 16 January 2006/ Accepted 24 February 2006

The hemA gene codes for one of two synthases in Rhodobacter sphaeroides 2.4.1 which catalyze the formation of 5-aminolevulinic acid. We have examined the role of PrrA, a DNA binding protein that is associated with the metabolic switch between aerobic growth and anoxygenic photosynthetic growth, in hemA expression and found that hemA transcription is directly activated by PrrA. Using electrophoretic mobility shift assays and DNase I protection assays, we have mapped two binding sites for PrrA within the hemA upstream sequences, each of which contains an identical 9-bp motif. Using lacZ transcription reporter plasmids in wild-type strain 2.4.1 and PrrA– mutant strain PRRA2, we showed that PrrA was required for maximal expression. We also found that the relative impacts of altering DNA sequences within the two binding sites are different depending on whether cells are growing aerobically or anaerobically. This reveals a greater level of complexity associated with PrrA-mediated regulation of transcription than has been heretofore described. Our findings are of particular importance with respect to those genes regulated by PrrA having more than one upstream binding site. In the case of the hemA gene, we discuss possibilities as to how these new insights can be accommodated within the context of what has already been established for hemA transcription regulation in R. sphaeroides.

* Corresponding author. Mailing address: Department of Biological Sciences, 374 Dodge Hall, Oakland University, Rochester, MI 48309. Phone: (248) 370-4497. Fax: (248) 370-4225. E-mail: zeilstra{at}oakland.edu.

Journal of Bacteriology, May 2006, p. 3208-3218, Vol. 188, No. 9
0021-9193/06/$08.00+0     doi:10.1128/JB.188.9.3208-3218.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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