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Journal of Bacteriology, December 2004, p. 8337-8346, Vol. 186, No. 24
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.24.8337-8346.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Bacillus subtilis Operon Encoding a Membrane Receptor for Bacteriophage SPP1{dagger}

Carlos São-José, Catarina Baptista, and Mário A. Santos*

Centro de Genética e Biologia Molecular e Departamento de Biologia Vegetal, Faculdade de Ciências da Universidade de Lisboa, Lisbon, Portugal

Received 28 June 2004/ Accepted 3 September 2004

The results reported here have identified yueB as the essential gene involved in irreversible binding of bacteriophage SPP1 to Bacillus subtilis. First, a deletion in an SPP1-resistant (pha-2) strain, covering most of the yueB gene, could be complemented by a xylose-inducible copy of yueB inserted at amyE. Second, disruption of yueB by insertion of a pMutin4 derivative resulted in a phage resistance phenotype regardless of the presence or absence of IPTG (isopropyl-ß-D-thiogalactopyranoside). YueB homologues are widely distributed in gram-positive bacteria. The protein Pip, which also serves as a phage receptor in Lactococcus lactis, belongs to the same family. yueB encodes a membrane protein of ~120 kDa, detected in immunoblots together with smaller forms that may be processed products arising from cleavage of its long extracellular domain. Insertional inactivation of yueB and the surrounding genes indicated that yueB is part of an operon which includes at least the upstream genes yukE, yukD, yukC, and yukBA. Disruption of each of the genes in the operon allowed efficient irreversible adsorption, provided that yueB expression was retained. Under these conditions, however, smaller plaques were produced, a phenotype which was particularly noticeable in yukE mutant strains. Interestingly, such reduction in plaque size was not correlated with a decreased adsorption rate. Overall, these results provide the first demonstration of a membrane-bound protein acting as a phage receptor in B. subtilis and suggest an additional involvement of the yukE operon in a step subsequent to irreversible adsorption.

* Corresponding author. Mailing address: Departamento de Biologia Vegetal, Faculdade de Ciências de Lisboa, Ed. ICAT, 1749-016 Lisbon, Portugal. Phone: (351) 217500000. Fax: (351)217500048. E-mail: mmsantos{at}fc.ul.pt.

{dagger} This paper is dedicated to Hermínia de Lencastre and Luís J. Archer, who first introduced M. A. Santos to the world of phage biology.

Journal of Bacteriology, December 2004, p. 8337-8346, Vol. 186, No. 24
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.24.8337-8346.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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