Nucleotide sequence of the cohesive single-stranded ends of Bacillus subtilis temperate bacteriophage phi 105

Abstract

The cohesive single-stranded ends of temperate Bacillus subtilis phage +105 were analyzed with the exonuclease activities of the Kienow fragment of DNA polymerase I and with exonuclease III and were found to be 3' extensions. Chemical sequencing of 3'-end-labeled fragments showed that the ends are 7-base extended 3' single strands and have the sequence: 5'-GCGCTCC-3'. 3'-CGCGAGG-5'

Mycobacteriophage Exploit NHEJ to Facilitate Genome Circularization

“…Indeed, comparison of sequences obtained directly from the linear genome with sequences obtained from the genome circularized by ligation confirmed that both Omega and Corndog have 4 base 3 0 complementary overhanging ends (Figure S6). In contrast, other lambda-like mycobacteriophage described in the same studies as Corndog and Omega (Pedulla et al, 2003), but lacking a phage Ku homolog, showed more typical nine or ten base overhangs expected to show a favorable association equilibrium (Ellis and Dean, 1985).…”

“…The ability of U-and Cd-Ku to facilitate LigD-dependent NHEJ predicted a role for NHEJ in the bacteriophage life cycle that, given that such a role was previously undocumented, in turn predicted that Omega and Corndog must have atypical features. Lambda-like dsDNA bacteriophage typically enter the cell as linear genomes, which are promptly circularized by self-association of long (R9 bases) complementary overhangs called cohesive (cos) ends (Ellis and Dean, 1985). We recently described the profile of rejoining of overhangs of varying length in yeast and showed that this length is a critical parameter in determining the repair mechanism, with repair becoming strictly Ku and NHEJ dependent when overhangs became shorter than six bases (Daley and Wilson, 2005).…”

Co-expression of a prophage system and a plasmid system in Bacillus subtilis

“…In this system, a plasmid system was used in combination with the 105MU331 prophage system to create a dual expression B. subtilis strain, in which the expression by the prophage system is heat inducible, whereas that of the plasmid system is constitutive. The 105MU331 prophage system [2] employed in this study was based on the B. subtilis phage 105 [3]. In this prophage system, a lacZ-cat cartridge is inserted into the lysogenic prophage, facilitating the integration of target genes into the host for the holin proteins [4] that cause lesions in the cytoplasmic membrane is deactivated by the insertion of the cloning cartridge [5].…”

Evidence for circular permutation of the prophage genome of Bacillus subtilis bacteriophage phi 105

“…Such colinearity is characteristic of coliphage Mu (17) and contrasts with the circular permutation exhibited by the prophages of coliphage lambda (5) and the B. subtilis bacteriophage SPO2 (6,14). Recently, the sequence of the 4105 cos site and the region surrounding it has been determined (9). No sequences similar to that of the lambda att site were found within the 4105 cos region, and unpublished data suggest that the 4105 att site is internally located (9).…”

“…Recently, the sequence of the 4105 cos site and the region surrounding it has been determined (9). No sequences similar to that of the lambda att site were found within the 4105 cos region, and unpublished data suggest that the 4105 att site is internally located (9). Recently, a detailed restriction map of the mature 4105 genome has been constructed (4).…”