Cryo-EM and cryo-ET reveal the molecular architecture and host interactions of mycobacteriophage Douge

Maharana, JitendraJitendraMaharanaWang, Chun-HsiungChun-HsiungWangTsai, Li-AnLi-AnTsaiLiao, Yi-TingYi-TingLiaoYang, Cheng-HanCheng-HanYangShen, Melvin C.Melvin C.ShenMacale, Lourriel S.Lourriel S.MacaleTran, Thang NgocThang NgocTranNarsico, JoemarkJoemarkNarsicoPerez, Ronelito J.Ronelito J.PerezTewary, Sunil KumarSunil KumarTewaryWu, Jian-LiJian-LiWuLin, Hong-YouHong-YouLinSHU-WEI CHANGFranklin, AaronAaronFranklinMoynihan, Patrick J.Patrick J.MoynihanJacobs-Sera, DeborahDeborahJacobs-SeraFreeman, Krista G.Krista G.FreemanHatfull, Graham F.Graham F.HatfullLowary, Todd L.Todd L.LowaryHo, Meng-ChiaoMeng-ChiaoHo2025-08-282025-08-282025-07-2426391856https://www.scopus.com/record/display.uri?eid=2-s2.0-105011273501&origin=resultslisthttps://scholars.lib.ntu.edu.tw/handle/123456789/731715Recent reports highlight the efficacy of engineered mycobacteriophages to treat non-tuberculosis mycobacterial disease. Molecular insights into mycobacteriophage architecture and host interactions could allow structure-guided phage engineering to increase efficacy and broaden host range, but such information is currently unavailable. We describe the cryoelectron microscopy (cryo-EM) structure of mycobacteriophage Douge, which contains 1,105 protein subunits assembled into a complete siphophage and is coated with glycan-binding domains for mycobacterial cell surface interactions. When filled with viral genome, the channel spanning the connector, tail, and baseplate is sealed by tape measure proteins, providing a genome gating system and requiring limited structural changes for genome ejection upon phage-host contact. Nanometer-resolution cryoelectron tomography (cryo-ET) snapshots of phage-host interactions show that the baseplate remains attached to the mycobacterial outer membrane during viral genome ejection. This study reveals high-resolution structural details of this mycobacteriophage and its interaction with host glycans.falseCP: MicrobiologyCP: Molecular biologycryo-EMcryo-ETlipoarabinomannan/arabinomannanmycobacteriophage structurephage-host interaction[SDGs]SDG2[SDGs]SDG3Cryo-EM and cryo-ET reveal the molecular architecture and host interactions of mycobacteriophage Dougejournal article10.1016/j.celrep.2025.1160572-s2.0-105011273501