Cloning of chlorophyllase, the key enzyme in chlorophyll degradation: Finding of a lipase motif and the induction by methyl jasmonate
- 21 December 1999
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 96 (26), 15362-15367
- https://doi.org/10.1073/pnas.96.26.15362
Abstract
Chlorophyllase (Chlase) is the first enzyme involved in chlorophyll (Chl) degradation and catalyzes the hydrolysis of ester bond to yield chlorophyllide and phytol. In the present study, we isolated the Chlase cDNA. We synthesized degenerate oligo DNA probes based on the internal amino acid sequences of purified Chlase from Chenopodium album, screened the C. album cDNA library, and cloned a cDNA (CaCLH, C. album chlorophyll-chlorophyllido hydrolase). The deduced amino acid sequence (347 aa residues) had a lipase motif overlapping with an ATP/GTP-binding motif (P-loop). CaCLH possibly was localized in the extraplastidic part of the cell, because a putative signal sequence for endoplasmic reticulum is at the N terminus. The amino acid sequence shared 37% identity with a function-unknown gene whose mRNA is inducible by coronatine and methyl jasmonate (MeJA) in Arabidopsis thaliana (AtCLH1). We expressed the gene products of AtCLH1 and of CaCLH in Escherichia coli, and they similarly exhibited Chlase activity. Moreover, we isolated another full-length cDNA based on an Arabidopsis genomic fragment and expressed it in E. coli, demonstrating the presence of the second Arabidopsis CLH gene (AtCLH2). No typical feature of signal sequence was identified in AtCLH1, whereas AtCLH2 had a typical signal sequence for chloroplast. AtCLH1 mRNA was induced rapidly by a treatment of MeJA, which is known to promote senescence and Chl degradation in plants, and a high mRNA level was maintained up to 9 h. AtCLH2, however, did not respond to MeJA.Keywords
This publication has 42 references indexed in Scilit:
- Chlorophyll: a symptom and a regulator of plastid developmentNew Phytologist, 1997
- Sequence Analysis of the Genome of the Unicellular Cyanobacterium Synechocystis sp. Strain PCC6803. II. Sequence Determination of the Entire Genome and Assignment of Potential Protein-coding RegionsDNA Research, 1996
- The Pseudomonas phytotoxin coronatine mimics octadecanoid signalling molecules of higher plantsFEBS Letters, 1994
- Arabidopsis Mutants Selected for Resistance to the Phytotoxin Coronatine Are Male Sterile, Insensitive to Methyl Jasmonate, and Resistant to a Bacterial PathogenTHE PLANT CELL ONLINE, 1994
- Directed Mutational Analysis of Bacteriochlorophyll a Biosynthesis in Rhodobacter capsulatusJournal of Molecular Biology, 1994
- Species‐specific variation in signal peptide design Implications for protein secretion in foreign hostsFEBS Letters, 1989
- Production of different pathogenic symptoms and different toxins by strains of Pseudomonas syringae pv. tomato not distinguishable by gel -immunodiffusion assayPhysiological Plant Pathology, 1983
- Identification of chlorophyllase as a glycoproteinFEBS Letters, 1981
- Properties of chloroplasts and chloroplast fragments, as deduced from internal chlorophyll → chlorophyllide conversionZeitschrift für Pflanzenphysiologie, 1974
- A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue CulturesPhysiologia Plantarum, 1962