Functional differentiation of two analogous coproporphyrinogen III oxidases for heme and chlorophyll biosynthesis pathways in the cyanobacterium Synechocystis sp. PCC 6803.
Coproporphyrinogen III oxidase (CPO) catalyzes the oxidative decarboxylation of coproporphyrinogen III to form protoporphyrinogen IX in heme biosynthesis and is shared in chlorophyll biosynthesis in photosynthetic organisms. There are two analogous CPOs, oxygen-dependent (HemF) and oxygen-independent (HemN) CPOs, in various organisms. Little information on cyanobacterial CPOs has been available to date. In the genome of the cyanobacterium Synechocystis sp. PCC 6803 there is one hemF-like gene, sll1185, and two hemN-like genes, sll1876 and sll1917. The three genes were overexpressed in Escherichia coli and purified to homogeneity. Sll1185 showed CPO activity under both aerobic and anaerobic conditions. While Sll1876 and Sll1917 showed absorbance spectra indicative of Fe-S proteins, only Sll1876 showed CPO activity under anaerobic conditions. Three mutants lacking one of these genes were isolated. The Deltasll1185 mutant failed to grow under aerobic conditions, with accumulation of coproporphyrin III. This growth defect was restored by cultivation under micro-oxic conditions. The growth of the Deltasll1876 mutant was significantly slower than that of the wild type under micro-oxic conditions, while it grew normally under aerobic conditions. Coproporphyrin III was accumulated at a low but significant level in the Deltasll1876 mutant grown under micro-oxic conditions. There was no detectable phenotype in Deltasll1917 under the conditions we examined. These results suggested that sll1185 encodes HemF as the sole CPO under aerobic conditions and that sll1876 encodes HemN operating under micro-oxic conditions, together with HemF. Such a differential operation of CPOs would ensure the stable supply of tetrapyrrole pigments under environments where oxygen levels fluctuate greatly.
Goto
Takeaki
T
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, 464-8601 Japan.
Aoki
Rina
R
Minamizaki
Kei
K
Fujita
Yuichi
Y
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002734:Chlorophyll / Q000096:biosynthesis*; D002851:Chromatography, High Pressure Liquid; D003304:Coproporphyrinogen Oxidase / Q000235:genetics / Q000378:metabolism*; D006418:Heme / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013050:Spectrometry, Fluorescence; D046939:Synechocystis / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D002734:Chlorophyll; D006418:Heme; D003304:Coproporphyrinogen Oxidase