Homotypic interactions among bacteriophage phiKMV early proteins.
Little is known about the bacteriophage proteins expressed immediately after infection of the host cell. Most of these early proteins are probably involved in bacteriophage-host interactions redirecting the bacterial metabolism to phage production. Interaction analysis of the first 16 phiKMV gene products (gp) identified homotypic interactions of gp7, gp9 and gp15. Two related yeast two-hybrid procedures, a matrix and a minilibrary approach, were applied to detect protein-protein interactions. A two-step selection procedure enabled drastic reduction of the background. Interactions were confirmed by drop tests. Multimerization of gp15 is consistent with its putative function as a DNA helicase involved in DNA replication. Homotypic interaction of gp7 and gp9 suggests they function as dimers or multimers. The absence of heterotypic interactions among early phiKMV proteins hints at their functional independence from other early phage proteins and their involvement in phage-host interactions that are important for creating optimal conditions for phage propagation. Besides, these results demonstrate the compatibility of phiKMV early gene products with the yeast two-hybrid system. Therefore, they are promising candidates to screen for interactions with host proteins.
Roucourt
B
B
Division of Gene Technology, Department of Biosystems, Katholieke Universiteit Leuven, Leuven, Belgium.
Chibeu
A
A
Lecoutere
E
E
Lavigne
R
R
Volckaert
G
G
Hertveldt
K
K
D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D019915:DNA Primase / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D019281:Dimerization; D017902:Podoviridae / Q000235:genetics / Q000378:metabolism*; D017105:Pseudomonas Phages / Q000235:genetics / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000821:virology; D020798:Two-Hybrid System Techniques; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism
D004268:DNA-Binding Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; D019915:DNA Primase; D004265:DNA Helicases