Characterization of an Amphipathic Alpha-Helix in the Membrane Targeting and Viral Genome Replication of Brome Mosaic Virus

Positive-strand RNA viruses associate with specific organelle membranes of host cells to establish viral replication complexes. The replication protein 1a of brome mosaic virus associates strongly with the nuclear endoplasmic reticulum (ER) membranes, invaginates membranes into the lumen, and recruits various host proteins to establish replication complexes termed spherules. 1a has a strong affinity towards the perinuclear ER membrane, however, the structural features in 1a that dictate its membrane associations and thereby membrane remodeling activities are unclear. This study examined the possible role of an amphipathic α-helix, helix B, in BMV 1a's membrane association. Deletion or single substitution of multiple amino acids of helix B abolished BMV 1a's localization to nuclear ER membranes. Additional reporter-based, gain-of-function assays showed that helix B is sufficient in targeting several soluble proteins to the nuclear ER membranes. Furthermore, we found that the helix B-mediated organelle targeting is a functionally conserved feature among positive-strand RNA viruses of the alphavirus-like superfamily that includes notable human viruses such as Hepatitis E virus and Rubella virus as well as plant viruses such as cucumber mosaic virus and cowpea chlorotic mottle virus. Our results demonstrate a critical role for helix B across members of the alphavirus-like superfamily in anchoring viral replication complexes to the organelle membranes. We anticipate our findings to be a starting point for the development of sophisticated models to use helix B as a novel target for the development of antivirals for positive-strand RNA viruses that belong to the alphavirus-like superfamily.

Name thesis:degree_name

Doctor of Philosophy

Level thesis:degree_level

doctoral

Discipline thesis:degree_discipline

Plant Pathology, Physiology and Weed Science

Department dc:contributor.department

Plant Pathology, Physiology and Weed Science

Grantor dc:publisher

Virginia Tech

Year dc:date.issued

2022

Author dc:creator

• Sathanantham, Preethi

Chair dc:contributor.committeechair

• Wang, Xiaofeng

Committee members dc:contributor.committeemember

• Capelluto, Daniel G.
• Meng, Xiang-Jin
• Vinatzer, Boris A.

• (+)RNA viruses
• alphavirus-like superfamily
• brome mosaic virus
• membrane associations
• amphipathic helices
• viral genome replication

Statement dc:rights

• In Copyright

Licence dc:rights.uri

• http://rightsstatements.org/vocab/InC/1.0/

Language dc:language.iso

en

Dc Identifier Other

vt_gsexam:33934

OAI identifier oai:identifier

oai:vtechworks.lib.vt.edu:10919/109007