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Journal of Bacteriology, August 2002, p. 4359-4368, Vol. 184, No. 16
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.16.4359-4368.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Genomic Analysis of Clostridium perfringens Bacteriophage {phi}3626, Which Integrates into guaA and Possibly Affects Sporulation

Markus Zimmer, Siegfried Scherer, and Martin J. Loessner*

Institut für Mikrobiologie, FML Weihenstephan, Technische Universität München, D-85350 Freising, Germany

Received 13 March 2002/ Accepted 17 May 2002

Two temperate viruses, {phi}3626 and {phi}8533, have been isolated from lysogenic Clostridium perfringens strains. Phage {phi}3626 was chosen for detailed analysis and was inspected by electron microscopy, protein profiling, and host range determination. For the first time, the nucleotide sequence of a bacteriophage infecting Clostridium species was determined. The virus belongs to the Siphoviridae family of the tailed phages, the order Caudovirales. Its genome consists of a linear double-stranded DNA molecule of 33,507 nucleotides, with invariable 3'-protruding cohesive ends of nine residues. Fifty open reading frames were identified, which are organized in three major life cycle-specific gene clusters. The genes required for lytic development show an opposite orientation and arrangement compared to the lysogeny control region. A function could be assigned to 19 gene products, based upon bioinformatic analyses, N-terminal amino acid sequencing, or experimental evidence. These include DNA-packaging proteins, structural components, a dual lysis system, a putative lysogeny switch, and proteins that are involved in replication, recombination, and modification of phage DNA. The presence of genes encoding a putative sigma factor related to sporulation-dependent sigma factors and a putative sporulation-dependent transcription regulator suggests a possible interaction of {phi}3626 with onset of sporulation in C. perfringens. We found that the {phi}3626 attachment site attP lies in a noncoding region immediately downstream of int. Integration of the viral genome occurs into the bacterial attachment site attB, which is located within the 3' end of a guaA homologue. This essential housekeeping gene is functionally independent of the integration status, due to reconstitution of its terminal codons by phage sequence.

* Corresponding author. Mailing address: Institut für Mikrobiologie, FML Weihenstephan, Technische Universität München, Weihenstephaner Berg 3, D-85350 Freising, Germany. Phone: 49 (8161) 71-3859. Fax: 49 (8161) 71-4492. E-mail: M.J.Loessner{at}Lrz.tum.de.

Journal of Bacteriology, August 2002, p. 4359-4368, Vol. 184, No. 16
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.16.4359-4368.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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