UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Publication Date

2020-02-03

Document Type

Article Postprint

Disciplines

Amino Acids, Peptides, and Proteins | Biochemistry | Molecular Biology | Structural Biology | Viruses

Abstract

Tailed bacteriophages use a DNA-packaging motor to encapsulate their genome during viral particle assembly. The small terminase (TerS) component of this DNA-packaging machinery acts as a molecular matchmaker that recognizes both the viral genome and the main motor component, the large terminase (TerL). However, how TerS binds DNA and the TerL protein remains unclear. Here, we identified gp83 of the thermophilic bacteriophage P74-26 as the TerS protein. We found that TerS(P76-26) oligomerizes into a nonamer that binds DNA, stimulates TerL ATPase activity, and inhibits TerL nuclease activity. A cryo-EM structure of TerS(P76-26) revealed that it forms a ring with a wide central pore and radially arrayed helix-turn-helix (HTH) domains. The structure further showed that these HTH domains, which are thought to bind DNA by wrapping the double helix around the ring, are rigidly held in an orientation distinct from that seen in other TerS proteins. This rigid arrangement of the putative DNA-binding domain imposed strong constraints on how TerS(P76-26) can bind DNA. Finally, the TerS(P76-26) structure lacked the conserved C-terminal beta-barrel domain used by other TerS proteins for binding TerL. This suggests that a well-ordered C-terminal beta-barrel domain is not required for TerS(P76-26) to carry out its matchmaking function. Our work highlights a thermophilic system for studying the role of small terminase proteins in viral maturation and presents the structure of TerS(P76-26), revealing key differences between this thermophilic phage and its mesophilic counterparts.

Keywords

DNA binding protein, DNA packaging, DNA recognition, bacteriophage, cryo-electron microscopy, helix-turn-helix domain, molecular motor, small terminase, thermophile, viral motor

Rights and Permissions

© 2020 The Author(s). Publisher's "Paper in Press" version posted as allowed by the publisher's author rights policy at https://www.asbmb.org/journals-news/editorial-policies.

DOI of Published Version

10.1074/jbc.RA119.012224

Source

J Biol Chem. 2020 Feb 3. pii: RA119.012224. doi: 10.1074/jbc.RA119.012224. Link to article on publisher's site

Journal/Book/Conference Title

The Journal of biological chemistry

Related Resources

Link to Article in PubMed

PubMed ID

32014998

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