The C-terminal domain of the bacteriophage T4 terminase docks on the prohead portal clip region during DNA packaging

Highlights

• A predicted structure for the T4 portal is proposed.

• Mutagenesis of portal clip region residues identified five residues unable to be substituted.

• The five residues include the proposed tunnel loop residue and all likely have important functions.

• FRET determined a 7.5 nm distance between clip residue 316 and terminase C-terminus.

• These studies support C-terminal nuclease domain docking of the terminase to the portal.

Abstract

Bacteriophage ATP-based packaging motors translocate DNA into a pre-formed prohead through a dodecameric portal ring channel to high density. We investigated portal–terminase docking interactions at specifically localized residues within a terminase-interaction region (aa279–316) in the phage T4 portal protein gp20 equated to the clip domain of the SPP1 portal crystal structure by 3D modeling. Within this region, three residues allowed A to C mutations whereas three others did not, consistent with informatics analyses showing the tolerated residues are not strongly conserved evolutionarily. About 7.5 nm was calculated by FCS-FRET studies employing maleimide Alexa488 dye labeled A316C proheads and gp17 CT-ReAsH supporting previous work docking the C-terminal end of the T4 terminase (gp17) closer to the N-terminal GFP-labeled portal (gp20) than the N-terminal end of the terminase. Such a terminase–portal orientation fits better to a proposed “DNA crunching” compression packaging motor and to portal determined DNA headful cutting.