Co-opting of nonATP-generating glycolytic enzymes for TBSV replication

Melissa Molho, Chingkai Chuang, Peter D. Nagy

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


Positive-strand RNA viruses build viral replication organelles (VROs) with the help of co-opted host factors. The energy requirement of intensive viral replication processes is less understood. Previous studies on tomato bushy stunt virus (TBSV) showed that tombusviruses hijack two ATP-producing glycolytic enzymes to produce ATP locally within VROs. In this work, we performed a cDNA library screen with Arabidopsis thaliana proteins and the TBSV p33 replication protein. The p33 - plant interactome contained highly conserved glycolytic proteins. We find that the glycolytic Hxk2 hexokinase, Eno2 phosphopyruvate hydratase and Fba1 fructose 1,6-bisphosphate aldolase are critical for TBSV replication in yeast or in a cell-free replicase reconstitution assay. The recruitment of Fba1 is important for the local production of ATP within VROs. Altogether, our data support the model that TBSV recruits and compartmentalizes possibly most members of the glycolytic pathway. This might allow TBSV to avoid competition with the host for ATP.

Original languageEnglish
Pages (from-to)15-29
Number of pages15
StatePublished - Jul 2021

Bibliographical note

Funding Information:
We thank Drs. Judit Pogany and Jun-ichi Inaba for comments on the manuscript. The authors thank Drs. H. Imamura and H. Noji for providing plasmids pRSET-ATeam YEMK and pRSET-ATeam RK . This work was supported by the National Science Foundation ( MCB-1517751 and IOS-1922895 ) and a USDA hatch grant ( KY012042 ) to PDN.

Publisher Copyright:
© 2021


  • ATP generation
  • ENO2
  • FBA1
  • Glycolysis
  • Hexokinase
  • Host factor
  • Replication
  • Tomato bushy stunt virus
  • Virus-host interaction
  • Yeast

ASJC Scopus subject areas

  • Virology


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