The chloroplast metalloproteases VAR2 and EGY1 act synergistically to regulate chloroplast development in Arabidopsis

J Biol Chem. 2020 Jan 24;295(4):1036-1046. doi: 10.1074/jbc.RA119.011853. Epub 2019 Dec 13.

Abstract

Chloroplast development and photosynthesis require the proper assembly and turnover of photosynthetic protein complexes. Chloroplasts harbor a repertoire of proteases to facilitate proteostasis and development. We have previously used an Arabidopsis leaf variegation mutant, yellow variegated2 (var2), defective in thylakoid FtsH protease complexes, as a tool to dissect the genetic regulation of chloroplast development. Here, we report a new genetic enhancer mutant of var2, enhancer of variegation3-1 (evr3-1). We confirm that EVR3 encodes a chloroplast metalloprotease, reported previously as ethylene-dependent gravitropism-deficient and yellow-green1 (EGY1)/ammonium overly sensitive1 (AMOS1). We observed that mutations in EVR3/EGY1/AMOS1 cause more severe leaf variegation in var2-5 and synthetic lethality in var2-4 Using a modified blue-native PAGE system, we reveal abnormal accumulations of photosystem I, photosystem II, and light-harvesting antenna complexes in EVR3/EGY1/AMOS1 mutants. Moreover, we discover distinct roles of VAR2 and EVR3/EGY1/AMOS1 in the turnover of photosystem II reaction center under high light stress. In summary, our findings indicate that two chloroplast metalloproteases, VAR2/AtFtsH2 and EVR3/EGY1/AMOS1, function coordinately to regulate chloroplast development and reveal new roles of EVR3/EGY1/AMOS1 in regulating chloroplast proteostasis in Arabidopsis.

Keywords: Arabidopsis thaliana; EGY1/AMOS1; VAR2/AtFtsH2; chloroplast; enhancer of variegation; leaf variegation; metalloprotease; photosynthesis; protein complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Dependent Proteases / genetics
  • ATP-Dependent Proteases / metabolism*
  • Amino Acid Sequence
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Base Sequence
  • Chloroplasts / metabolism*
  • Etiolation
  • Genetic Loci
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Metalloproteases / metabolism*
  • Mutation / genetics
  • Photosynthesis
  • Photosystem I Protein Complex / metabolism
  • Photosystem II Protein Complex / metabolism
  • Protein Multimerization
  • Protein Stability

Substances

  • Arabidopsis Proteins
  • Membrane Proteins
  • Photosystem I Protein Complex
  • Photosystem II Protein Complex
  • EGY1 protein, Arabidopsis
  • Metalloproteases
  • ATP-Dependent Proteases
  • VAR2 protein, Arabidopsis