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Journal of Bacteriology, June 2004, p. 3525-3530, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3525-3530.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Structure of the DNA-SspC Complex: Implications for DNA Packaging, Protection, and Repair in Bacterial Spores

Daphna Frenkiel-Krispin,1 Rinat Sack,1 Joseph Englander,1 Eyal Shimoni,1 Miriam Eisenstein,2 Esther Bullitt,3 Rachel Horowitz-Scherer,4 Christopher S. Hayes,5,{dagger} Peter Setlow,5 Abraham Minsky,1 and Sharon Grayer Wolf2*

Organic Chemistry,1 Chemical Research Support, The Weizmann Institute of Science, Rehovot 76100, Israel,2 Department of Physiology & Biophysics, Boston University School of Medicine, Boston, Massachusetts 02118,3 Department of Biology, University of Massachusetts, Amherst, Massachusetts 01003,4 Department of Biochemistry, University of Connecticut Health Center, Farmington, Connecticut 060325

Received 21 December 2003/ Accepted 20 February 2004

Bacterial spores have long been recognized as the sturdiest known life forms on earth, revealing extraordinary resistance to a broad range of environmental assaults. A family of highly conserved spore-specific DNA-binding proteins, termed {alpha}/ß-type small, acid-soluble spore proteins (SASP), plays a major role in mediating spore resistance. The mechanism by which these proteins exert their protective activity remains poorly understood, in part due to the lack of structural data on the DNA-SASP complex. By using cryoelectron microscopy, we have determined the structure of the helical complex formed between DNA and SspC, a characteristic member of the {alpha}/ß-type SASP family. The protein is found to fully coat the DNA, forming distinct protruding domains, and to modify DNA structure such that it adopts a 3.2-nm pitch. The protruding SspC motifs allow for interdigitation of adjacent DNA-SspC filaments into a tightly packed assembly of nucleoprotein helices. By effectively sequestering DNA molecules, this dense assembly of filaments is proposed to enhance and complement DNA protection obtained by DNA saturation with the {alpha}/ß-type SASP.

* Corresponding author. Mailing address: Chemical Research Support, The Weizmann Institute of Science, Rehovot 76100, Israel. Phone: 972-8-934-4421. Fax: 972-8-934-4161. E-mail: Sharon.Wolf{at}weizmann.ac.il.

{dagger} Present address: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.

Journal of Bacteriology, June 2004, p. 3525-3530, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3525-3530.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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