Functional analysis of N-terminal domains of Arabidopsis chlorophyllide a oxygenase.
Higher plants acclimate to various light environments by changing the antenna size of a light-harvesting photosystem. The antenna size of a photosystem is partly determined by the amount of chlorophyll b in the light-harvesting complexes. Chlorophyllide a oxygenase (CAO) converts chlorophyll a to chlorophyll b in a two-step oxygenation reaction. In our previous study, we demonstrated that the cellular level of the CAO protein controls accumulation of chlorophyll b. We found that the amino acids sequences of CAO in higher plants consist of three domains (A, B, and C domains). The C domain exhibits a catalytic function, and we demonstrated that the combination of the A and B domains regulates the cellular level of CAO. However, the individual function of each of A and B domain has not been determined yet. Therefore, in the present study we constructed a series of deleted CAO sequences that were fused with green fluorescent protein and overexpressed in a chlorophyll b-less mutant of Arabidopsis thaliana, ch1-1, to further dissect functions of A and B domains. Subsequent comparative analyses of the transgenic plants overexpressing B domain containing proteins and those lacking the B domain determined that there was no significant difference in CAO protein levels. These results indicate that the B domain is not involved in the regulation of the CAO protein levels. Taken together, we concluded that the A domain alone is involved in the regulatory mechanism of the CAO protein levels.
Sakuraba
Yasuhito
Y
Institute of Low Temperature Science, Hokkaido University, N19 W8, Sapporo, 060-0819, Japan. sakuraba@pop.lowtem.hokudai.ac.jp
Yamasato
Akihiro
A
Tanaka
Ryouichi
R
Tanaka
Ayumi
A
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D002734:Chlorophyll / Q000378:metabolism; D002735:Chlorophyllides / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting; D018613:Microscopy, Confocal; D008954:Models, Biological; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D017386:Sequence Homology, Amino Acid
D029681:Arabidopsis Proteins; D002735:Chlorophyllides; D002734:Chlorophyll; D049452:Green Fluorescent Proteins; C034388:chlorophyllide a; C037184:chlorophyll b; D010105:Oxygenases