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J Biol Chem ; 276(23): 19999-20004, 2001 Jun 08.
Article in English | MEDLINE | ID: mdl-11264284

ABSTRACT

Synechocystis PCC 6803 contains four genes encoding polypeptides with sequence features of CPx-type ATPases, two of which are now designated pacS and ctaA. We show that CtaA and PacS (but not the related transporters, ZiaA or CoaT) facilitate switching to the use of copper (in plastocyanin) as an alternative to iron (in cytochrome c(6)) for the carriage of electrons within the thylakoid lumen. Disruption of pacS reduced copper tolerance but enhanced silver tolerance, and pacS-mediated restoration of copper tolerance was used to select transformants. Disruption of ctaA caused no change in copper tolerance but reduced the amount of copper cell(-1). In cultures supplemented with 0.2 microm copper, photooxidation of cytochrome c(6) (PetJ) was depressed in wild-type cells but remained elevated in both Synechocystis PCC 6803(ctaA) and Synechocystis PCC 6803(pacS). Conversely, plastocyanin transcripts (petE) were less abundant in both mutants at this [copper]. Synechocystis PCC 6803(ctaA) and Synechocystis PCC 6803(pacS) showed increased iron dependence with impaired growth in deferoxamine mesylate (iron chelator)-containing media. Double mutants also deficient in cytochrome c(6), Synechocystis PCC 6803(petJ,ctaA) and Synechocystis PCC 6803(petJ,pacS), were viable, but the former had increased copper dependence with severely impaired growth in the presence of bathocuproinedisulfonic acid (copper chelator). Analogous transporters are likely to supply copper to plastocyanin in chloroplasts.


Subject(s)
Adenosine Triphosphatases/metabolism , Carrier Proteins/metabolism , Cation Transport Proteins , Copper/metabolism , Cyanobacteria/physiology , Photosynthesis , Recombinant Fusion Proteins , Base Sequence , Cyanobacteria/enzymology , Cyanobacteria/genetics , Cytochromes/genetics , Cytochromes f , DNA Primers , Genes, Bacterial , Mutation , Plastocyanin/genetics , RNA, Messenger/genetics
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