Differential expression of two isopentenyl pyrophosphate isomerases and enhanced carotenoid accumulation in a unicellular chlorophyte.
The enzyme isopentenyl pyrophosphate (IPP) isomerase catalyzes the reversible isomerization of IPP to produce dimethylallyl pyrophosphate, the initial substrate leading to the biosynthesis of carotenoids and many other long-chain isoprenoids. Expression of IPP isomerase, and of two enzymes specific to the carotenoid pathway (lycopene beta-cyclase and beta-carotene-C-4-oxygenase), was followed in the green unicellular alga Haematococcus pluvialis after exposure to high illumination. This alga uniquely accumulates carotenoids in the cytoplasm and in late developmental stages turns deep-red in color because of accumulation of ketocarotenoids in the cytosol. The carotenoid/chlorophyll ratio increased 3-fold in wild type and 6-fold in a precocious carotenoid-accumulating mutant (Car-3) within 24 h after increasing the illumination from 20 to 150 micromol photon m-2.s-1. Two cDNAs encoding IPP isomerase in Haematococcus, ipiHp1 and ipiHp2, were identified. Although otherwise highly similar (95% identity overall), the predicted sequence of ipiHp1 contained a 12-aa region not found in that of ipiHp2. This was reflected by a size difference between two polypeptides of 34 and 32.5 kDa, both of which reacted with an antibody to the product of ipiHp1. We suggest that the 32.5-kDa form is involved with the carotenoid accumulation in the cytoplasm, since the 32.5-kDa polypeptide was preferentially up-regulated by high light preceding the carotenoid increase and only this form was detected in red cysts.
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Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA.
Cunningham
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Gantt
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D000595:Amino Acid Sequence; D000818:Animals; D019748:Carbon-Carbon Double Bond Isomerases / Q000737:chemistry*; D002338:Carotenoids / Q000096:biosynthesis*; D002734:Chlorophyll / Q000378:metabolism; D000460:Chlorophyta / Q000201:enzymology*; D003001:Cloning, Molecular; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D045782:Hemiterpenes; D019753:Intramolecular Lyases / Q000032:analysis; D007527:Isoenzymes / Q000378:metabolism; D008027:Light; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D009154:Mutation / Q000235:genetics; D009943:Organophosphorus Compounds / Q000378:metabolism; D010860:Pigments, Biological / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment; D017421:Sequence Analysis
D045782:Hemiterpenes; D007527:Isoenzymes; D009943:Organophosphorus Compounds; D010860:Pigments, Biological; D012333:RNA, Messenger; D002734:Chlorophyll; C004809:isopentenyl pyrophosphate; D002338:Carotenoids; D019748:Carbon-Carbon Double Bond Isomerases; C021453:isopentenyldiphosphate delta-isomerase; D019753:Intramolecular Lyases; C070925:lycopene cyclase-isomerase