title: Phage-borne factors and host LexA regulate the lytic switch in phage GIL01.
abstract: The Bacillus thuringiensis temperate phage GIL01 does not integrate into the host chromosome but exists stably as an independent linear replicon within the cell. Similar to that of the lambdoid prophages, the lytic cycle of GIL01 is induced as part of the cellular SOS response to DNA damage. However, no CI-like maintenance repressor has been detected in the phage genome, suggesting that GIL01 uses a novel mechanism to maintain lysogeny. To gain insights into the GIL01 regulatory circuit, we isolated and characterized a set of 17 clear plaque (cp) mutants that are unable to lysogenize. Two phage-encoded proteins, gp1 and gp7, are required for stable lysogen formation. Analysis of cp mutants also identified a 14-bp palindromic dinBox1 sequence within the P1-P2 promoter region that resembles the known LexA-binding site of Gram-positive bacteria. Mutations at conserved positions in dinBox1 result in a cp phenotype. Genomic analysis identified a total of three dinBox sites within GIL01 promoter regions. To investigate the possibility that the host LexA regulates GIL01, phage induction was measured in a host carrying a noncleavable lexA (Ind(-)) mutation. GIL01 formed stable lysogens in this host, but lytic growth could not be induced by treatment with mitomycin C. Also, mitomycin C induced β-galactosidase expression from GIL01-lacZ promoter fusions, and induction was similarly blocked in the lexA (Ind(-)) mutant host. These data support a model in which host LexA binds to dinBox sequences in GIL01, repressing phage gene expression during lysogeny and providing the switch necessary to enter lytic development.
authors: Laboratory of Food and Environmental Microbiology, Universite´ Catholique de Louvain, Croix du Sud, 2, Box L7.05.12,B-1348 Louvain-la-Neuve, Belgium.
authors: Nadine
authors: N
authors: Fornelos
authors: Jaana K H
authors: JK
authors: Bamford
authors: Jacques
authors: J
authors: Mahillon
mesh_terms: D017103:Bacillus Phages / Q000235:genetics / Q000254:growth & development*; D001413:Bacillus thuringiensis / Q000235:genetics / Q000821:virology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001433:Bacteriolysis; D001665:Binding Sites; D004279:DNA, Viral / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D006790:Host-Parasite Interactions; D008242:Lysogeny; D011485:Protein Binding; D013014:SOS Response, Genetics; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D014775:Virus Activation; D000099160:LexA Repressor Protein
chemical_list: D001426:Bacterial Proteins; D004279:DNA, Viral; D012697:Serine Endopeptidases; D000099160:LexA Repressor Protein