Analysis of the Bacillus subtilis Bacteriophages SPP1 and SF6 Gene 1 Product: A Protein Involved in the Initiation of Headful Packaging

Abstract

Gene 1 product (G1P) of the related Bacillus subtilis bacteriophages SPP1, SF6, and ρ15 is essential for DNA maturation and packaging. A DNA segment containing gene 1 of phage SF6 or ρ15 origin was cloned and sequenced. SF6 and ρ15 G1P (both with predicted molecular mass of 16.7 kDa) share 71% identity with G1P of SPP1. The G1P of all three phages contains three conserved segments (I, II, and III). Within segments I and II helix-turn-helix DNA binding and nucleotide binding motifs were predicted. G1P of both SPP1 and SF6 origin was purified. SPP1 G1P protein (20.7 kDa), purified from cells overexpressing the cloned gene, purifies together with another polypeptide, having a molecular mass of about 13 kDa. The 13-kDa polypeptide results from a translation start signal within gene 1, and hence was termed SPP1 G1P*. G1P of both SPP1 and SF6 binds specifically to a pac -containing DNA fragment, whereas G1P*, which lacks segment I, does not. Chimeric G1P protiens were obtained by domain swapping between gene 1 of SPP1 and SF6. The results presented here suggest that the G1P DNA binding motif lies in segment I and the major determinant for G1P::G1P interaction might lie in segment II. Segment III and the extended C-terminal part of SPP1 G1P are dispensable. The G1P::G2P interacting region remains uncharacterized.