Nuclear mutants of Chlamydomonas reinhardtii defective in the biogenesis of the cytochrome b6f complex.
The random integration of transforming DNA into the nuclear genome of Chlamydomonas has been employed as an insertional mutagen to generate a collection of photosynthetic mutants that display abnormal steady-state fluorescence levels and an acetate-requiring phenotype. Electron paramagnetic resonance spectroscopy was then used to identify those mutants that specifically lack a functional cytochrome b6f complex. Our analysis of RNA and protein synthesis in five of these mutants reveals four separate phenotypes. One mutant fails to accumulate transcript for cytochrome f, whilst a second displays a severely reduced accumulation of the cytochrome b6 transcript. Two other mutants appear to be affected in the insertion of the haem co-factor into cytochrome b6. The fifth mutant displays no detectable defect in the synthesis of any of the known subunits of the complex. Genetic analysis of the mutants demonstrates that in three cases, the mutant phenotype co-segregates with the introduced DNA. For the mutant affected in the accumulation of the cytochrome f transcript, we have used the introduced DNA as a tag to isolate the wild-type version of the affected gene.
Gumpel
N J
NJ
Department of Biology, University College London, UK.
Ralley
L
L
Girard-Bascou
J
J
Wollman
F A
FA
Nugent
J H
JH
Purton
S
S
D000818:Animals; D001483:Base Sequence; D016825:Chlamydomonas reinhardtii / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D003573:Cytochrome b Group / Q000096:biosynthesis*; D045346:Cytochrome b6f Complex; D008040:Genetic Linkage; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010788:Photosynthesis / Q000235:genetics; D010957:Plasmids
D003573:Cytochrome b Group; D045346:Cytochrome b6f Complex