An Arabidopsis thaliana virescent mutant reveals a role for ClpR1 in plastid development

• Published in: Plant molecular biology 2007-01, Vol.63 (1), p.85-96
• Main Authors: Koussevitzky, Shai, Stanne, Tara M, Peto, Charles A, Giap, Tony, Sjögren, Lars L E, Zhao, Yunde, Clarke, Adrian K, Chory, Joanne
• Format: Article
• Language: English
• Subjects: Arabidopsis - genetics; Arabidopsis - physiology; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Arabidopsis Proteins - physiology; Arabidopsis thaliana; ATP; ATP-DEPENDENT PROTEASE; Biodegradation; Biological Sciences; Biologiska vetenskaper; Blotting, Northern; Blotting, Western; chloroplast development; Chloroplasts; Chloroplasts - genetics; Chloroplasts - physiology; Chloroplasts - ultrastructure; Clp protease; DEGRADATION; E coli; ENCODES; Endopeptidase Clp - genetics; Endopeptidase Clp - metabolism; Endopeptidase Clp - physiology; ESCHERICHIA-COLI; FTSH; GENE; Gene Expression Regulation, Developmental; Gene Expression Regulation, Plant; LEAF VARIEGATION; Microscopy, Electron, Transmission; MOLECULAR CHAPERONE; Mutation; Plant Leaves - growth & development; Plant Leaves - metabolism; Plants, Genetically Modified; Plastids - genetics; Plastids - physiology; Plastids - ultrastructure; Proteins; RIBOSOMAL-RNAS; RNA, Plant - genetics; RNA, Plant - metabolism; RNA, Ribosomal, 23S - genetics; RNA, Ribosomal, 23S - metabolism; virescent
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1007/s11103-006-9074-2

The ATP-dependent Clp protease has been well-characterized in Escherichia coli, but knowledge of its function in higher plants is limited. In bacteria, this two-component protease consists of a Ser-type endopeptidase ClpP, which relies on the ATP-dependent unfolding activity from an Hsp100 molecular chaperone to initiate protein degradation. In the chloroplasts of higher plants, multiple isoforms of the proteolytic subunit exist, with Arabidopsis having five ClpPs and four ClpP-like proteins termed ClpR predicted in its genome. In this work we characterized an Arabidopsis mutant impaired in one subunit of the chloroplast-localized Clp protease core, ClpR1. clpR1-1, a virescent mutant, carries a pre-mature stop codon in the clpR1 gene, resulting in no detectable ClpR1 protein. The accumulation of several chloroplast proteins, as well as most of the chloroplast-localized Clp protease subunits, is inhibited in clpR1-1. Unexpectedly, some plastid-encoded proteins do not accumulate, although their transcripts accumulate to wild-type levels. Maturation of 23S and 4.5S chloroplast ribosomal RNA (cp-rRNA) is delayed in clpR1-1, and both RNAs accumulate as higher molecular weight precursors. Also, chloroplasts in clpR1-1 are smaller than in wild type and have fewer thylakoid membranes with smaller grana stacks. We propose that a ClpR1-containing activity is required for chloroplast development and differentiation and in its absence both are delayed.