Portal de Periódicos da CAPES

Directed chloroplast transformation in Chlamydomonas reinhardtii: insertional inactivation of the psaC gene encoding the iron sulfur protein destabilizes photosystem I.

1991; Springer Nature; Volume: 10; Issue: 8 Linguagem: Inglês

10.1002/j.1460-2075.1991.tb07733.x

1460-2075

Yuichiro Takahashi, Michel Goldschmidt‐Clermont, Satoshi Soen, Lars‐Gunnar Franzén, Jean‐David Rochaix,

Metalloenzymes and iron-sulfur proteins

Research Article1 August 1991free access Directed chloroplast transformation in Chlamydomonas reinhardtii: insertional inactivation of the psaC gene encoding the iron sulfur protein destabilizes photosystem I. Y. Takahashi Y. Takahashi Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author M. Goldschmidt-Clermont M. Goldschmidt-Clermont Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author S.Y. Soen S.Y. Soen Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author L.G. Franzén L.G. Franzén Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author J.D. Rochaix J.D. Rochaix Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author Y. Takahashi Y. Takahashi Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author M. Goldschmidt-Clermont M. Goldschmidt-Clermont Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author S.Y. Soen S.Y. Soen Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author L.G. Franzén L.G. Franzén Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author J.D. Rochaix J.D. Rochaix Department of Molecular Biology, University of Geneva, Switzerland. Search for more papers by this author Author Information Y. Takahashi1, M. Goldschmidt-Clermont1, S.Y. Soen1, L.G. Franzén1 and J.D. Rochaix1 1Department of Molecular Biology, University of Geneva, Switzerland. The EMBO Journal (1991)10:2033-2040https://doi.org/10.1002/j.1460-2075.1991.tb07733.x PDFDownload PDF of article text and main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info The chloroplast gene psaC encoding the iron sulfur protein of photosystem I (PSI) from the green alga Chlamydomonas reinhardtii has been cloned and characterized. The deduced amino acid sequence is highly related to that of higher plants and cyanobacteria. Using a particle gun, wild type C. reinhardtii cells have been transformed with a plasmid carrying the psaC gene disrupted by an aadA gene cassette designed to express spectinomycin/streptomycin resistance in the chloroplast. Transformants selected on plates containing acetate as a reduced carbon source and spectinomycin are unable to grow on minimal medium lacking acetate and are deficient in PSI activity. Southern blot analysis of total cell DNA of the transformants shows that the wild type psaC gene has been replaced by the interrupted psaC gene through homologous recombination. While authentic transcripts of the psaC gene are no longer detected, aadA gives rise to a few transcripts in the transformants. Biochemical analysis indicates that neither PSI reaction center subunits nor the seven small subunits belonging to PSI accumulate stably in the thylakoid membranes of the transformants. Pulse-chase labeling of cell proteins shows that the PSI reaction center subunits are synthesized normally but turn over rapidly in the transformants. We conclude that the iron sulfur binding protein encoded by the psaC gene is an essential component, both for photochemical activity and for stable assembly of PSI. The present study suggests that any chloroplast gene encoding a component of the photosynthetic apparatus can be disrupted in C. reinhardtii using the strategy described. Previous ArticleNext Article Volume 10Issue 81 August 1991In this issue RelatedDetailsLoading ...