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Journal of Bacteriology, December 2008, p. 8204-8214, Vol. 190, No. 24
0021-9193/08/$08.00+0     doi:10.1128/JB.00752-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Deletion of cgR_1596 and cgR_2070, Encoding NlpC/P60 Proteins, Causes a Defect in Cell Separation in Corynebacterium glutamicum R{triangledown} ,{dagger}

Yota Tsuge,1,2,{ddagger},§ Hidetaka Ogino,1,§ Haruhiko Teramoto,1 Masayuki Inui,1 and Hideaki Yukawa1,2*

Research Institute of Innovative Technology for the Earth, 9-2 Kizugawadai, Kizugawa, Kyoto 619-0292, Japan,1 Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama-cho, Ikoma, Nara 630-0101, Japan2

Received 27 May 2008/ Accepted 1 October 2008

In previous work, random genome deletion mutants of Corynebacterium glutamicum R were generated using the insertion sequence (IS) element IS31831 and the Cre/loxP excision system. One of these mutants, C. glutamicum strain RD41, resulting from the deletion of a 10.1-kb genomic region ({Delta}cgR_1595 through cgR_1604) from the WT strain, showed cell elongation, and several lines appeared on the cell surface (bamboo shape). The morphological changes were suppressed by overexpression of cgR_1596. Single disruption of cgR_1596 in WT C. glutamicum R resulted in morphological changes similar to those observed in the RD41 strain. CgR_1596 has a predicted secretion signal peptide in the amino-terminal region and a predicted NlpC/P60 domain, which is conserved in cell wall hydrolases, in the carboxyl-terminal region. In C. glutamicum R, CgR_0802, CgR_1596, CgR_2069, and CgR_2070 have the NlpC/P60 domain; however, only simultaneous disruption of cgR_1596 and cgR_2070, and not cgR_2070 single disruption, resulted in cell growth delay and more severe morphological changes than disruption of cgR_1596. Transmission electron microscopy revealed multiple septa within individual cells of cgR_1596 single and cgR_1596-cgR_2070 double disruptants. Taken together, these results suggest that cgR_1596 and cgR_2070 are involved in cell separation and cell growth in C. glutamicum.

* Corresponding author. Mailing address: Research Institute of Innovative Technology for the Earth, 9-2 Kizugawadai, Kizugawa, Kyoto 619-0292, Japan. Phone: (81) 774 75 2308. Fax: (81) 774 75 2321. E-mail: mmg-lab{at}rite.or.jp

{triangledown} Published ahead of print on 17 October 2008.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

{ddagger} Present address: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520.

§ These authors contributed equally to this work.

Journal of Bacteriology, December 2008, p. 8204-8214, Vol. 190, No. 24
0021-9193/08/$08.00+0     doi:10.1128/JB.00752-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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