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Journal of Bacteriology, August 2001, p. 4786-4795, Vol. 183, No. 16
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.16.4786-4795.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Genetic Studies of mrp, a Locus Essential for Cellular Aggregation and Sporulation of Myxococcus xanthus

Hong Sun and Wenyuan Shi*

Molecular Biology Institute and School of Dentistry, University of California, Los Angeles, California 90095

Received 9 April 2001/Accepted 25 May 2001

Under starvation conditions, Myxococcus xanthus undergoes a complex developmental process which includes cellular aggregation and sporulation. A transposon insertion mutant (the Tn5-Omega 280 mutant) with defects in both aggregation and sporulation was analyzed in this study. The Tn5-Omega 280 mutant was found to have a disrupted NtrC-like response regulator designated Myxococcus regulatory protein B (mrpB). Further sequencing analyses revealed a histidine kinase homolog (mrpA) immediately upstream of mrpB and a cyclic AMP receptor protein-like transcriptional regulator (mrpC) downstream of mrpB. In-frame deletion analyses revealed that both the mrpB and mrpC genes were required for cellular aggregation and sporulation but that only mrpA was required for sporulation only. Site-specific mutagenesis of the putative phosphorylation site of MrpB, D58, showed that a D58A mutation caused defects in both aggregation and sporulation but that a D58E mutation resulted in only a sporulation defect. Further genetic and molecular analyses with reporter genes and reverse transcription-PCR indicated that mrpA and mrpB are cotranscribed but that mrpC is transcribed independently and that all of these genes are developmentally regulated. In addition, MrpB is essential for transcription of mrpC and MrpC regulates its own transcription. These data indicate that Mrp proteins are important components required for M. xanthus development. The complicated interaction between Mrp proteins may play an important role in regulating developmental gene expression in M. xanthus.

* Corresponding author. Mailing address: UCLA Molecular Biology Institute and School of Dentistry, P.O. Box 951668, 10833 Le Conte Ave., Los Angeles, CA 90095-1668. Phone: (310) 825-8356. Fax: (310) 794-7109. E-mail: wenyuan{at}ucla.edu.

Journal of Bacteriology, August 2001, p. 4786-4795, Vol. 183, No. 16
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.16.4786-4795.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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