Coordinated regulation and complex formation of yellow variegated1 and yellow variegated2, chloroplastic FtsH metalloproteases involved in the repair cycle of photosystem II in Arabidopsis thylakoid membranes.
Arabidopsis yellow variegated1 (VAR1) and VAR2 are separate loci that encode similar chloroplast FtsH proteases. To date, FtsH is the best-characterized protease in thylakoid membranes involved in the turnover of photosynthetic protein complexes. It comprises a protein family that is encoded by 12 different nuclear genes in Arabidopsis. We show here that nine FtsH proteins are located in the chloroplasts. Mutations in either VAR1 or VAR2 cause typical leaf variegation and sensitivity to photoinhibition. By contrast, none of these phenotypes was observed in T-DNA insertion mutants in other ftsH genes (ftsh1, ftsh6, and ftsh8) closely related to VAR1 and VAR2. This finding suggests that VAR1 and VAR2 play a predominant role in the photosystem II repair cycle in thylakoid membranes. By generating VAR1- and VAR2-specific antibodies, we found that loss of either VAR1 or VAR2 results in the decreased accumulation of the other. Thus, the genetic nonredundancy between VAR1 and VAR2 could be attributed to their coordinated regulation at the protein level. These observations led us to examine whether VAR1 and VAR2 form a complex. Sucrose density gradient and gel filtration analyses revealed a complex of approximately 400 to 450 kD, probably representing a hexamer. Furthermore, VAR1 and VAR2 were shown to coprecipitate by immunoprecipitation using VAR1- and VAR2-specific antibodies. The majority of VAR1 appears to exist as heterocomplexes with VAR2, whereas VAR2 may be present as homocomplexes as well. Based on these results, we conclude that VAR1 and VAR2 are the major components of an FtsH complex involved in the repair of photodamaged proteins in thylakoid membranes.
Sakamoto
Wataru
W
Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan. saka@rib.okayama-u.ac.jp
Zaltsman
Adi
A
Adam
Zach
Z
Takahashi
Yuichiro
Y
D049069:ATP-Dependent Proteases; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D008027:Light; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D045726:Metalloproteases / Q000235:genetics* / Q000378:metabolism; D016254:Mutagenesis, Insertional; D009154:Mutation; D010641:Phenotype; D045332:Photosystem II Protein Complex / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D020524:Thylakoids / Q000201:enzymology / Q000235:genetics* / Q000528:radiation effects
D049069:ATP-Dependent Proteases; D029681:Arabidopsis Proteins; D045643:DNA Repair Enzymes; D004269:DNA, Bacterial; D008565:Membrane Proteins; D045726:Metalloproteases; D045332:Photosystem II Protein Complex; C036483:T-DNA; C402651:AT2G30950 protein, Arabidopsis