Helper component-proteinase enhances the activity of 1-deoxy-D-xylulose-5-phosphate synthase and promotes the biosynthesis of plastidic isoprenoids in Potato virus Y-infected tobacco

Plant Cell Environ. 2015 Oct;38(10):2023-34. doi: 10.1111/pce.12526. Epub 2015 Apr 27.

Abstract

Virus-infected plants show strong morphological and physiological alterations. Many physiological processes in chloroplast are affected, including the plastidic isoprenoid biosynthetic pathway [the 2C-methyl-D-erythritol-4-phosphate (MEP) pathway]; indeed, isoprenoid contents have been demonstrated to be altered in virus-infected plants. In this study, we found that the levels of photosynthetic pigments and abscisic acid (ABA) were altered in Potato virus Y (PVY)-infected tobacco. Using yeast two-hybrid assays, we demonstrated an interaction between virus protein PVY helper component-proteinase (HC-Pro) and tobacco chloroplast protein 1-deoxy-D-xylulose-5-phosphate synthase (NtDXS). This interaction was confirmed using bimolecular fluorescence complementation (BiFC) assays and pull-down assays. The Transket_pyr domain (residues 394-561) of NtDXS was required for interaction with HC-Pro, while the N-terminal region of HC-Pro (residues 1-97) was necessary for interaction with NtDXS. Using in vitro enzyme activity assays, PVY HC-Pro was found to promote the synthase activity of NtDXS. We observed increases in photosynthetic pigment contents and ABA levels in transgenic plants with HC-Pro accumulating in the chloroplasts. During virus infection, the enhancement of plastidic isoprenoid biosynthesis was attributed to the enhancement of DXS activity by HC-Pro. Our study reveals a new role of HC-Pro in the host plant metabolic system and will contribute to the study of host-virus relationships.

Keywords: ABA; DXS; MEP pathway; photosynthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abscisic Acid / metabolism
  • Base Sequence
  • Chloroplast Proteins / genetics
  • Chloroplast Proteins / metabolism
  • Chloroplasts / enzymology
  • Cysteine Endopeptidases / metabolism*
  • Gene Library
  • Molecular Sequence Data
  • Nicotiana / enzymology*
  • Nicotiana / genetics
  • Nicotiana / metabolism
  • Nicotiana / virology
  • Photosynthesis
  • Plant Diseases / virology*
  • Plant Growth Regulators / metabolism
  • Plant Leaves / enzymology
  • Plant Leaves / genetics
  • Plant Leaves / metabolism
  • Plant Leaves / virology
  • Plants, Genetically Modified
  • Plastids / metabolism
  • Potyvirus / physiology*
  • Sequence Analysis, DNA
  • Sequence Deletion
  • Terpenes / metabolism*
  • Transferases / genetics
  • Transferases / metabolism*
  • Two-Hybrid System Techniques
  • Viral Proteins / metabolism*

Substances

  • Chloroplast Proteins
  • Plant Growth Regulators
  • Terpenes
  • Viral Proteins
  • Abscisic Acid
  • Transferases
  • deoxyxylulose-5-phosphate synthase
  • Cysteine Endopeptidases
  • HC-Pro protein, potyvirus

Associated data

  • GENBANK/AAD38941
  • GENBANK/AF143812
  • GENBANK/EU650419
  • GENBANK/O78328
  • GENBANK/X02353