Assembly-associated structural changes of bacteriophage T7 capsids. Detection by use of a protein-specific probe.
To detect changes in capsid structure that occur when a preassembled bacteriophage T7 capsid both packages and cleaves to mature-size longer (concatameric) DNA, the kinetics and thermodynamics are determined here for the binding of the protein-specific probe, 1,1'-bi(4-anilino)naphthalene-5,5'-di-sulfonic acid (bis-ANS), to bacteriophage T7, a T7 DNA deletion (8.4%) mutant, and a DNA-free T7 capsid (metrizamide low density capsid II) known to be a DNA packaging intermediate that has a permeability barrier not present in a related capsid (metrizamide high density capsid II). Initially, some binding to either bacteriophage or metrizamide low density capsid II occurs too rapidly to quantify (phase 1, duration < 10 s). Subsequent binding (phase 2) occurs with first-order kinetics. Only the phase 1 binding occurs for metrizamide high density capsid II. These observations, together with both the kinetics of the quenching by ethidium of bound bis-ANS fluorescence and the nature of bis-ANS-induced protein alterations, are explained by the hypothesis that the phase 2 binding occurs at internal sites. The number of these internal sites increases as the density of the packaged DNA decreases. The accompanying change in structure is potentially the signal for initiating cleavage of a concatemer. Evidence for the following was also obtained: (a) a previously undetected packaging-associated change in the conformation of the major protein of the outer capsid shell and (b) partitioning by a permeability barrier of the interior of the T7 capsid.
Khan
S A
SA
Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284.
Griess
G A
GA
Serwer
P
P
D000817:Anilino Naphthalenesulfonates; D055592:Biophysical Phenomena; D001703:Biophysics; D002213:Capsid / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes; D007700:Kinetics; D015394:Molecular Structure; D013050:Spectrometry, Fluorescence; D013604:T-Phages / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D013816:Thermodynamics
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate)