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Originally published In Press as doi:10.1074/jbc.M111214200 on May 7, 2002

J. Biol. Chem., Vol. 277, Issue 30, 26753-26760, July 26, 2002
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Role of fruA and csgA Genes in Gene Expression during Development of Myxococcus xanthus
ANALYSIS BY TWO-DIMENSIONAL GEL ELECTROPHORESIS*,

Takayuki HoriuchiDagger §, Masato Taoka, Toshiaki Isobe, Teruya KomanoDagger ||, and Sumiko Inouye§**

From the Departments of Dagger  Biology and  Chemistry, Tokyo Metropolitan University, Minamiohsawa, Hachioji, Tokyo 192-0397, Japan and the § Department of Biochemistry, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854

Two genes, fruA and csgA, encoding a putative transcription factor and C-factor, respectively, are essential for fruiting body formation of Myxococcus xanthus. To investigate the role of fruA and csgA genes in developmental gene expression, developing cells as well as vegetative cells of M. xanthus wild-type, fruA::Tc, and csgA731 strains were pulse-labeled with [35S]methionine, and the whole cell proteins were analyzed using two-dimensional immobilized pH gradient/SDS-PAGE. Differences in protein synthesis patterns among more than 700 protein spots were detected during development of the three strains. Fourteen proteins showing distinctly different expression patterns in mutant cells were analyzed in more detail. Five of the 14 proteins were identified as elongation factor Tu (EF-Tu), Dru, DofA, FruA, and protein S by immunoblot analysis and mass spectroscopy. A gene encoding DofA was cloned and sequenced. Although both fruA and csgA genes regulate early development of M. xanthus, they were found to differently regulate expression of several developmental genes. The production of six proteins, including DofA and protein S, was dependent on fruA, whereas the production of two proteins was dependent on csgA, and one protein was dependent on both fruA and csgA. To explain the present findings, a new model was presented in which different levels of FruA phosphorylation may distinctively regulate the expression of two groups of developmental genes.

* This work was supported in part by a grant from the Ministry of Education, Science, Sports and Culture of Japan and by the Foundation of Medicine and Dentistry of New Jersey.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains the results of a detailed analysis of Fig. 1.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AB073986 and AB073987.

|| To whom correspondence may be addressed. Tel.: 81-426-77-2568; Fax: 81-426-77-2559; E-mail: komano-teruya@c.metro-u.ac.jp.

** To whom correspondence may be addressed: Dept. of Biochemistry, Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, NJ 08854. Tel.: 732-235-4161; Fax: 732-235-4783, E-mail: sinouye@waksman.rutgers.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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