Abstract
The I subunit of magnesium-chelatase (CHLI) is encoded by two genes in Arabidopsis (Arabidopsis thaliana), CHLI1 and CHLI2. Conflicting results have been reported concerning the functions of the two proteins. We show here that the chli1/chli1 chli2/chli2 double knockout mutant was albino. Comparison with the pale-green phenotype of a chli1/chli1 single knockout mutant indicates that CHLI2 could support some chlorophyll biosynthesis in the complete absence of CHLI1. Real-time quantitative reverse transcription-polymerase chain reaction showed that CHLI2 was expressed at a much lower level than CHLI1. The chli1/chli1 chli2/chli2 double mutant could be fully rescued by expressing a transgene of CHLI2 driven by the CHLI1 promoter. These results suggest that differences between CHLI1 and CHLI2 lie mostly in their expression levels. Furthermore, both the chli1/chli1 and chli2/chli2 single knockout mutants had lower survival rates during de-etiolation than the wild type, suggesting that both genes are required for optimal growth during de-etiolation. In addition, we show that a semidominant chli1 mutant allele and the chli1/chli1 chli2/chli2 double mutant accumulated Lhcb1 transcripts when treated with the herbicide norflurazon, indicating that knocking out the CHLI activity causes the genome-uncoupled phenotype.
MeSH Terms
Amino Acid Sequence
Arabidopsis/enzymology,genetics
Arabidopsis Proteins/chemistry,genetics,metabolism
Chlorophyll/biosynthesis
Gene Expression Regulation, Plant
Immunoblotting
Lyases/chemistry,genetics,metabolism
Molecular Sequence Data
Mutation/genetics
Phenotype
Promoter Regions, Genetic/genetics
Protein Stability
RNA, Messenger/genetics,metabolism
Chemicals
Arabidopsis Proteins
RNA, Messenger
Chlorophyll
Lyases
magnesium chelatase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Huang Yi-Shiuan
Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan.
Li Hsou-Min
References (25)
25 references, click to expand
-
GUN4, a regulator of chlorophyll synthesis and intracellular signaling.
Science. 2003 Feb 7;299(5608):902-6
PMID: 12574634
-
Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays.
Nat Biotechnol. 2000 Jun;18(6):630-4
PMID: 10835600
-
Plastid-to-nucleus retrograde signaling.
Annu Rev Plant Biol. 2006;57:739-59
PMID: 16669780
-
Gene silencing from plant DNA carried by a Geminivirus.
Plant J. 1998 Apr;14(1):91-100
PMID: 15494056
-
Three semidominant barley mutants with single amino acid substitutions in the smallest magnesium chelatase subunit form defective AAA+ hexamers.
Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13944-9
PMID: 12357035
-
Functional analysis of Arabidopsis thaliana isoforms of the Mg-chelatase CHLI subunit.
Photochem Photobiol Sci. 2008 Oct;7(10):1188-95
PMID: 18846282
-
Magnesium chelatase: association with ribosomes and mutant complementation studies identify barley subunit Xantha-G as a functional counterpart of Rhodobacter subunit BchD.
Mol Gen Genet. 1997 Mar 18;254(1):85-92
PMID: 9108294
-
Tetrapyrrole profiling in Arabidopsis seedlings reveals that retrograde plastid nuclear signaling is not due to Mg-protoporphyrin IX accumulation.
Proc Natl Acad Sci U S A. 2008 Sep 30;105(39):15178-83
PMID: 18818314
-
An Arabidopsis mutant that is resistant to the protoporphyrinogen oxidase inhibitor acifluorfen shows regulatory changes in tetrapyrrole biosynthesis.
Mol Genet Genomics. 2005 Jun;273(4):311-8
PMID: 15815918
-
The steady-state level of Mg-protoporphyrin IX is not a determinant of plastid-to-nucleus signaling in Arabidopsis.
Proc Natl Acad Sci U S A. 2008 Sep 30;105(39):15184-9
PMID: 18818313
-
Gene expression profiling of the tetrapyrrole metabolic pathway in Arabidopsis with a mini-array system.
Plant Physiol. 2004 Aug;135(4):2379-91
PMID: 15326282
-
Magnesium-protoporphyrin chelatase of Rhodobacter sphaeroides: reconstitution of activity by combining the products of the bchH, -I, and -D genes expressed in Escherichia coli.
Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):1941-4
PMID: 7892204
-
Analysis of gun phenotype in barley magnesium chelatase and Mg-protoporphyrin IX monomethyl ester cyclase mutants.
Plant Physiol Biochem. 2005 Oct-Nov;43(10-11):901-8
PMID: 16310365
-
Floral spray transformation can efficiently generate Arabidopsis transgenic plants.
Transgenic Res. 2000 Dec;9(6):471-6
PMID: 11206976
-
In vivo visualization of Mg-protoporphyrin IX, a coordinator of photosynthetic gene expression in the nucleus and the chloroplast.
Plant Cell. 2007 Jun;19(6):1964-79
PMID: 17586657
-
Arabidopsis genomes uncoupled 5 (GUN5) mutant reveals the involvement of Mg-chelatase H subunit in plastid-to-nucleus signal transduction.
Proc Natl Acad Sci U S A. 2001 Feb 13;98(4):2053-8
PMID: 11172074
-
Signal transduction mutants of Arabidopsis uncouple nuclear CAB and RBCS gene expression from chloroplast development.
Cell. 1993 Sep 10;74(5):787-99
PMID: 7690685
-
Three separate proteins constitute the magnesium chelatase of Rhodobacter sphaeroides.
Eur J Biochem. 1996 Jan 15;235(1-2):438-43
PMID: 8631364
-
The small, versatile pPZP family of Agrobacterium binary vectors for plant transformation.
Plant Mol Biol. 1994 Sep;25(6):989-94
PMID: 7919218
-
Molecular basis for semidominance of missense mutations in the XANTHA-H (42-kDa) subunit of magnesium chelatase.
Proc Natl Acad Sci U S A. 1999 Feb 16;96(4):1744-9
PMID: 9990095
-
Chlorophyll biosynthesis. Expression of a second chl I gene of magnesium chelatase in Arabidopsis supports only limited chlorophyll synthesis.
Plant Physiol. 2002 Feb;128(2):770-9
PMID: 11842180
-
GENEVESTIGATOR. Arabidopsis microarray database and analysis toolbox.
Plant Physiol. 2004 Sep;136(1):2621-32
PMID: 15375207
-
Chloroplast to nucleus communication triggered by accumulation of Mg-protoporphyrinIX.
Nature. 2003 Jan 2;421(6918):79-83
PMID: 12511958
-
The analysis of the ChlI 1 and ChlI 2 genes using acifluorfen-resistant mutant of Arabidopsis thaliana.
Planta. 2007 Mar;225(4):935-43
PMID: 17123104
-
Transposon tagging of the sulfur gene of tobacco using engineered maize Ac/Ds elements.
Genetics. 1999 Dec;153(4):1919-28
PMID: 10581296