Dimerization between the holin and holin inhibitor of phage lambda.
Holins are integral membrane proteins that control the access of phage-encoded muralytic enzymes, or endolysins, to the cell wall by the sudden formation of an uncharacterized homo-oligomeric lesion, or hole, in the membrane, at a precisely defined time. The timing of lambda-infected cell lysis depends solely on the 107 codon S gene, which encodes two proteins, S105 and S107, which are the holin and holin inhibitor, respectively. Here we report the results of biochemical and genetic studies on the interaction between the holin and the holin inhibitor. A unique cysteine at position 51, in the middle of the second transmembrane domain, is shown to cause the formation of disulfide-linked dimers during detergent membrane extraction. Forced oxidation of membranes containing S molecules also results in the formation of covalently linked dimers. This technique is used to demonstrate efficient dimeric interactions between S105 and S107. These results, coupled with the previous finding that the timing of lysis depends on the excess of the amount of S105 over S107, suggest a model in which the inhibitor functions by titrating out the effector in a stoichiometric fashion. This provides a basis for understanding two evolutionary advantages provided by the inhibitor system, in which the production of the inhibitor not only causes a delay in the timing of lysis, allowing the assembly of more virions, but also increases effective hole formation after triggering.
Gründling
A
A
Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843-2128, USA.
Smith
D L
DL
Bläsi
U
U
Young
R
R
D001433:Bacteriolysis; D010582:Bacteriophage lambda / Q000737:chemistry / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D003902:Detergents; D019281:Dimerization; D004220:Disulfides; D013997:Time Factors; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics
D003902:Detergents; D004220:Disulfides; D014764:Viral Proteins; C082001:bacteriophage lambda lysis effector protein S105; C082002:bacteriophage lambda lysis inhibitor protein S107; D003545:Cysteine