title: Probing the structure of the S105 hole.
abstract: For most phages, holins control the timing of host lysis. During the morphogenesis period of the infection cycle, canonical holins accumulate harmlessly in the cytoplasmic membrane until they suddenly trigger to form lethal lesions called holes. The holes can be visualized by cryo-electron microscopy and tomography as micrometer-scale interruptions in the membrane. To explore the fine structure of the holes formed by the lambda holin, S105, a cysteine-scanning accessibility study was performed. A collection of S105 alleles encoding holins with a single Cys residue in different positions was developed and characterized for lytic function. Based on the ability of 4-acetamido-4'-((iodoacetyl) amino) stilbene-2,2'-disulfonic acid, disodium salt (IASD), to modify these Cys residues, one face of transmembrane domain 1 (TMD1) and TMD3 was judged to face the lumen of the S105 hole. In both cases, the lumen-accessible face was found to correspond to the more hydrophilic face of the two TMDs. Judging by the efficiency of IASD modification, it was concluded that the bulk of the S105 protein molecules were involved in facing the lumen. These results are consistent with a model in which the perimeters of the S105 holes are lined by the holin molecules present at the time of lysis. Moreover, the findings that TMD1 and TMD3 face the lumen, coupled with previous results showing TMD2-TMD2 contacts in the S105 dimer, support a model in which membrane depolarization drives the transition of S105 from homotypic to heterotypic oligomeric interactions.
authors.affiliation: Center for Phage Technology, Department of Biochemistry and Biophysics, and Department of Biology, Texas A&M University, College Station, Texas, USA.
authors.forename: Kam H
authors.initials: KH
authors.lastname: To
authors.affiliation: Center for Phage Technology, Department of Biochemistry and Biophysics, and Department of Biology, Texas A&M University, College Station, Texas, USA ryland@tamu.edu.
authors.forename: Ry
authors.initials: R
authors.lastname: Young
mesh_terms: D000595:Amino Acid Sequence; D010582:Bacteriophage lambda / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000821:virology*; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D008969:Molecular Sequence Data; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D057074:Virus Release / Q000502:physiology
chemical_list: D014764:Viral Proteins; C082001:bacteriophage lambda lysis effector protein S105
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