Structural Analysis and Expression during Dark-Light Transitions of a Gene for Cytochrome f in Chlamydomonas reinhardtii1

Author(s):  
Luis R. Comolli ◽  
Jianhui Zhou ◽  
Thomas Linden ◽  
Rainer Breitling ◽  
Jorge Flores ◽  
...  

1988 ◽  
Vol 43 (3-4) ◽  
pp. 207-212 ◽  
Author(s):  
Roland Pschorn ◽  
Wolfgang Rühle ◽  
Aloysius Wild

Ferredoxin-NADP+-oxidoreductase (FNR, EC 1.18.1.2) has been shown to be activated by light within a few seconds during dark-light transitions and inactivated in the dark. In previous papers this could be pointed out by the correlation of cytochrome f induction kinetics to the rate of NADP-photoreduction and the variable fluorescence. The present study deals with the role of the proton gradient during the activation process. The transition from an inactive to an active form is followed continuously in an in situ system. The steady-state rate of NADP-photoreduction is affected only by ionophores which inhibit a formation of the proton gradient, but not by inhibitors of the electric field. It correlates to the 9-aminoacridine fluorescence quench and the light scattering signals at 535 nm. The perception of the pH-gradient through the enzyme is still a matter of discussion.


1994 ◽  
Vol 14 (9) ◽  
pp. 6171-6179 ◽  
Author(s):  
N R Sturm ◽  
R Kuras ◽  
S Büschlen ◽  
W Sakamoto ◽  
K L Kindle ◽  
...  

FUD6, a nonphotosynthetic mutant of Chlamydomonas reinhardtii, was previously found to be deficient in the synthesis of subunit IV of the cytochrome b6/f complex, the chloroplast petD gene product (C. Lemaire, J. Girard-Bascou, F.-A. Wollman, and P. Bennoun, Biochim. Biophys. Acta 851:229-238, 1986). The lesion in FUD6 is a 236-bp deletion between two 11-bp direct repeats in the chloroplast genome. It extends from 82 to 72 bp upstream of the 5' end of wild-type petD mRNA to 156 to 166 bp downstream of the 5' end. Thus, the deletion extends into the putative promoter and 5' untranslated region of petD. No petD mRNA of the normal size can be detected in FUD6 cells, but a low level of a dicistronic message accumulates, which contains the coding regions for subunit IV and cytochrome f, the product of the upstream petA gene. petD transcriptional activity in FUD6 is not significantly altered from the wild-type level. This transcriptional activity was eliminated by petA promoter disruptions, suggesting that it originates at the petA promoter. We conclude that the petD-coding portion of most cotranscripts is rapidly degraded in FUD6, possibly following processing events that generate the 3' end of petA mRNA. A chloroplast transformant was constructed in which only the sequence from -81 to -2 relative to the major 5' end of the petD transcript was deleted. Although this deletion eliminates all detectable petD promoter activity, the transformant grows phototrophically and accumulates high levels of monocistronic petD mRNA. We conclude that the petD gene can be transcribed by functionally redundant promoters. In the absence of a functional petD promoter, a lack of transcription termination allows the downstream petD gene to be cotranscribed with the petA coding region and thereby expressed efficiently.


2007 ◽  
Vol 342 (17) ◽  
pp. 2557-2566 ◽  
Author(s):  
Klaus Bollig ◽  
Marc Lamshöft ◽  
Kristian Schweimer ◽  
Franz-Josef Marner ◽  
Herbert Budzikiewicz ◽  
...  

2006 ◽  
Vol 275 (4) ◽  
pp. 387-398 ◽  
Author(s):  
Loreta Gudynaite-Savitch ◽  
Michael Gretes ◽  
Rachael M. Morgan-Kiss ◽  
Leonid V. Savitch ◽  
John Simmonds ◽  
...  

Genetics ◽  
2001 ◽  
Vol 159 (1) ◽  
pp. 119-132
Author(s):  
K Wostrikoff ◽  
Y Choquet ◽  
F-A Wollman ◽  
J Girard-Bascou

Abstract We isolated seven allelic nuclear mutants of Chlamydomonas reinhardtii specifically blocked in the translation of cytochrome f, a major chloroplast-encoded subunit of the photosynthetic electron transport chain encoded by the petA gene. We recovered one chloroplast suppressor in which the coding region of petA was now expressed under the control of a duplicated 5′ untranslated region from another open reading frame of presently unknown function. Since we also recovered 14 nuclear intragenic suppressors, we ended up with 21 alleles of a single nuclear gene we called TCA1 for translation of cytochrome b6f complex petA mRNA. The high number of TCA1 alleles, together with the absence of genetic evidence for other nuclear loci controlling translation of the chloroplast petA gene, strongly suggests that TCA1 is the only trans-acting factor. We studied the assembly-dependent regulation of cytochrome f translation—known as the CES process—in TCA1-mutated contexts. In the presence of a leaky tca1 allele, we observed that the regulation of cytochrome f translation was now exerted within the limits of the restricted translational activation conferred by the altered version of TCA1 as predicted if TCA1 was the ternary effector involved in the CES process.


Biochemistry ◽  
1995 ◽  
Vol 34 (22) ◽  
pp. 7468-7475 ◽  
Author(s):  
Richard Kuras ◽  
Francis-Andre Wollman ◽  
Pierre Joliot

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