An isoform of eukaryotic initiation factor 4E from Chrysanthemum morifolium interacts with Chrysanthemum virus B coat protein

PLoS One. 2013;8(3):e57229. doi: 10.1371/journal.pone.0057229. Epub 2013 Mar 7.

Abstract

Background: Eukaryotic translation initiation factor 4E (eIF4E) plays an important role in plant virus infection as well as the regulation of gene translation.

Methodology/principal findings: Here, we describe the isolation of a cDNA encoding CmeIF(iso)4E (GenBank accession no. JQ904592), an isoform of eIF4E from chrysanthemum, using RACE PCR. We used the CmeIF(iso)4E cDNA for expression profiling and to analyze the interaction between CmeIF(iso)4E and the Chrysanthemum virus B coat protein (CVBCP). Multiple sequence alignment and phylogenetic tree analysis showed that the sequence similarity of CmeIF(iso)4E with other reported plant eIF(iso)4E sequences varied between 69.12% and 89.18%, indicating that CmeIF(iso)4E belongs to the eIF(iso)4E subfamily of the eIF4E family. CmeIF(iso)4E was present in all chrysanthemum organs, but was particularly abundant in the roots and flowers. Confocal microscopy showed that a transiently transfected CmeIF(iso)4E-GFP fusion protein distributed throughout the whole cell in onion epidermis cells. A yeast two hybrid assay showed CVBCP interacted with CmeIF(iso)4E but not with CmeIF4E. BiFC assay further demonstrated the interaction between CmeIF(iso)4E and CVBCP. Luminescence assay showed that CVBCP increased the RLU of Luc-CVB, suggesting CVBCP might participate in the translation of viral proteins.

Conclusions/significance: These results inferred that CmeIF(iso)4E as the cap-binding subunit eIF(iso)4F may be involved in Chrysanthemum Virus B infection in chrysanthemum through its interaction with CVBCP in spatial.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Capsid Proteins / metabolism*
  • Chrysanthemum / genetics
  • Chrysanthemum / metabolism*
  • Chrysanthemum / virology*
  • Eukaryotic Initiation Factor-4E / classification
  • Eukaryotic Initiation Factor-4E / genetics
  • Eukaryotic Initiation Factor-4E / metabolism*
  • Gene Expression Profiling
  • Molecular Sequence Data
  • Phylogeny
  • Plant Viruses / metabolism*
  • Protein Binding
  • Protein Biosynthesis
  • Protein Isoforms / classification
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • RNA, Viral / genetics
  • RNA, Viral / metabolism

Substances

  • Capsid Proteins
  • Eukaryotic Initiation Factor-4E
  • Protein Isoforms
  • RNA, Viral

Associated data

  • GENBANK/JQ904592

Grants and funding

This study is supported by the Program for New Century Excellent Talents in University of Chinese Ministry of Education (Grant No. NCET-10-0492), Sci-Tech Support Plan of Jiangsu Province (BE2011325), 863 Project (2011AA100208), a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) and the Fundamental Research Funds for the Central Universities (KYZ201147) and Youth Science and Technology Innovation Fund (KJ2011009). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.