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The Effect of Two Amino acid Residue Substitutions via RNA Editing on Dark-operative Protochlorophyllide Oxidoreductase in the Black Pine Chloroplasts.
- Source :
-
Scientific reports [Sci Rep] 2017 May 24; Vol. 7 (1), pp. 2377. Date of Electronic Publication: 2017 May 24. - Publication Year :
- 2017
-
Abstract
- Dark-operative protochlorophyllide oxidoreductase (DPOR) is a key enzyme to produce chlorophyll in the dark. Among photosynthetic eukaryotes, all three subunits chlL, chlN, and chlB are encoded by plastid genomes. In some gymnosperms, two codons of chlB mRNA are changed by RNA editing to codons encoding evolutionarily conserved amino acid residues. However, the effect of these substitutions on DPOR activity remains unknown. We first prepared cyanobacterial ChlB variants with amino acid substitution(s) to mimic ChlB translated from pre-edited mRNA. Their activities were evaluated by measuring chlorophyll content of dark-grown transformants of a chlB-lacking mutant of the cyanobacterium Leptolyngbya boryana that was complemented with pre-edited mimic chlB variants. The chlorophyll content of the transformant cells expressing the ChlB variant from the fully pre-edited mRNA was only one-fourth of the control cells. Co-purification experiments of ChlB with Strep-ChlN suggested that a stable complex with ChlN is greatly impaired in the substituted ChlB variant. We then confirmed that RNA editing efficiency was markedly greater in the dark than in the light in cotyledons of the black pine Pinus thunbergii. These results indicate that RNA editing on chlB mRNA is important to maintain appropriate DPOR activity in black pine chloroplasts.
- Subjects :
- Amino Acid Sequence
Amino Acid Substitution
Bacterial Proteins genetics
Bacterial Proteins metabolism
Chloroplasts genetics
Cloning, Molecular
Cyanobacteria genetics
Darkness
Escherichia coli genetics
Escherichia coli metabolism
Gene Expression
Genetic Vectors chemistry
Genetic Vectors metabolism
Models, Molecular
Oxidoreductases Acting on CH-CH Group Donors genetics
Oxidoreductases Acting on CH-CH Group Donors metabolism
Photosynthesis genetics
Pinus genetics
Plant Proteins genetics
Plant Proteins metabolism
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protochlorophyllide metabolism
RNA Editing
RNA, Messenger genetics
RNA, Messenger metabolism
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins metabolism
Sequence Alignment
Structural Homology, Protein
Bacterial Proteins chemistry
Chloroplasts enzymology
Cyanobacteria enzymology
Oxidoreductases Acting on CH-CH Group Donors chemistry
Pinus enzymology
Plant Proteins chemistry
Protochlorophyllide chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 7
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 28539650
- Full Text :
- https://doi.org/10.1038/s41598-017-02630-2