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Lettuce Big-Vein Associated Virus ORF3 Encodes a Functional 30K Movement Protein.

Willem E W Schravesande1,2, Machiel V Cligge1, Raoul Frijters2

  • 1Molecular Plant Pathology, Swammerdam Institute for Life Sciences (SILS), University of Amsterdam, Amsterdam, the Netherlands.

Molecular Plant Pathology
|September 25, 2025
PubMed
Summary

Lettuce big-vein associated virus ORF3 acts as a movement protein, facilitating plant virus spread by localizing to plasmodesmata. This discovery aids in understanding viral movement and disease development.

Keywords:
lettucelettuce big‐vein diseasemovement proteinplasmodesmataprotein structure

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Area of Science:

  • Plant virology
  • Molecular biology
  • Structural biology

Background:

  • Movement proteins (MPs) are crucial for plant virus cell-to-cell and systemic movement.
  • The 30K superfamily represents the largest group of MPs, characterized by conserved jelly-roll structures despite low sequence similarity.
  • Lettuce big-vein associated virus (LBVaV) is linked to severe lettuce big-vein disease (LBVD) and facilitates the spread of other viruses, but its proteins remain uncharacterized.

Purpose of the Study:

  • To identify and characterize the movement proteins of Lettuce big-vein associated virus (LBVaV).
  • To investigate the role of LBVaV ORF3 in plant virus movement and its potential classification within the 30K superfamily of MPs.

Main Methods:

  • Utilized AlphaFold2 for protein structure prediction and FoldSeek for structure-based homology searches to annotate LBVaV open reading frames (ORFs).
  • Performed microscopy to determine the subcellular localization of LBVaV ORF3.
  • Conducted in planta expression studies to assess the ability of ORF3 to facilitate the movement of movement-impaired viruses.

Main Results:

  • All LBVaV ORFs were annotated, with ORF3 identified as belonging to the 30K superfamily of MPs.
  • LBVaV ORF3 exhibits low sequence identity (5-11%) to related MPs but shares the characteristic jelly-roll fold.
  • Microscopy confirmed ORF3 localization at plasmodesmata, and its expression enhanced cell-to-cell movement of two distinct plant viruses.

Conclusions:

  • The AlphaFold2-FoldSeek strategy successfully identified a novel MP (LBVaV ORF3) despite low sequence homology.
  • LBVaV ORF3 functions as a movement protein, localizing to plasmodesmata and promoting viral spread.
  • This finding contributes to understanding the diversity of MPs and viral movement mechanisms in plants.