DNA macroarray and real-time PCR analysis of two nuclear photosystem I mutants from Chlamydomonas reinhardtii reveal downregulation of Lhcb genes but different regulation of Lhca genes

Balczun, Carsten; Bunse, Astrid; Nowrousian, Minou; Korbel, A.; Glanz, Stephanie; Kück, Ulrich

doi:10.1016/j.bbaexp.2005.11.006

Cited by 5 publications

(2 citation statements)

References 38 publications

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“…Five micrograms of total RNA were reverse‐transcribed by SuperScript III Reverse Transcriptase (Invitrogen), according to the manufacturer's instructions. Gene‐specific primers were designed to amplify fragments of approximately 100–350 bp in length for the C. reinhardtii dxs , dxr , ipi , ggps , psy , pds , lcyb , and bchy genes in the carotenoid metabolic pathway, for CrFtsZ1 (Wang et al 2003) and CrOr (Lu et al 2006) for plastid development and carotenoid accumulation, and for actin (Sangiorgio et al 2004; Yehudai‐Resheff et al 2007), α‐tubulin (Balczun et al 2005), RACK1 (Mus et al 2007), and ypt C1 (Lake and Willows 2003) as candidate reference genes. A list of genes, GenBank accession numbers, and forward and reverse primers is provided in Table 1.…”

Section: Methodsmentioning

confidence: 99%

Coordinated Regulation of Gene Expression for Carotenoid Metabolism in Chlamydomonas reinhardtii

Sun

Liu

Hui

et al. 2010

JIPB

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Carotenoids are important plant pigments for both light harvesting and photooxidation protection. Using the model system of the unicellular green alga Chlamydomonas reinhardtii, we characterized the regulation of gene expression for carotenoid metabolism by quantifying changes in the transcript abundance of dxs, dxr and ipi in the plastidic methylerythritol phosphate pathway and of ggps, psy, pds, lcyb and bchy, directly involved in carotenoid metabolism, under different photoperiod, light and metabolite treatments. The expression of these genes fluctuated with light/dark shifting. Light treatment also promoted the accumulation of transcripts of all these genes. Of the genes studied, dxs, ggps and lcyb displayed the typical circadian pattern by retaining a rhythmic fluctuation of transcript abundance under both constant light and constant dark entrainments. The expression of these genes could also be regulated by metabolic intermediates. For example, ggps was significantly suppressed by a geranylgeranyl pyrophosphate supplement and ipi was upregulated by isopentenyl pyrophosphate. Furthermore, CrOr, a C. reinhardtii homolog of the recently characterized Or gene that accounts for carotenoid accumulation, also showed co-expression with carotenoid biosynthetic genes such as pds and lcyb. Our data suggest a coordinated regulation on carotenoid metabolism in C. reinhardtii at the transcriptional level.

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Section: Methodsmentioning

confidence: 99%

Coordinated Regulation of Gene Expression for Carotenoid Metabolism in Chlamydomonas reinhardtii

Sun

Liu

Hui

et al. 2010

JIPB

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“…Escherichia coli strain XL1-blue MRF′ served as host for general plasmid construction and maintenance ( 19 ). The C.reinhardtii cDNA library contains double-stranded cDNA fragments with EcoRI adaptors cloned into the EcoRI site of pGAD10 in DH10B (BD Biosciences Clontech, Heidelberg, Germany) ( 20 ). pGAD10 derivatives were used in the yeast three-hybrid screen.…”

Section: Methodsmentioning

confidence: 99%

A nucleosome assembly protein-like polypeptide binds to chloroplast group II intron RNA in Chlamydomonas reinhardtii

Glanz

Bunse²,

Wimbert³

et al. 2006

Nucleic Acids Research

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In the unicellular green alga Chlamydomonas reinhardtii, the chloroplast-encoded tscA RNA is part of a tripartite group IIB intron, which is involved in trans-splicing of precursor mRNAs. We have used the yeast three-hybrid system to identify chloroplast group II intron RNA-binding proteins, capable of interacting with the tscA RNA. Of 14 candidate cDNAs, 13 encode identical polypeptides with significant homology to members of the nuclear nucleosome assembly protein (NAP) family. The RNA-binding property of the identified polypeptide was demonstrated by electrophoretic mobility shift assays using different domains of the tripartite group II intron as well as further chloroplast transcripts. Because of its binding to chloroplast RNA it was designated as NAP-like (cNAPL). In silico analysis revealed that the derived polypeptide carries a 46 amino acid chloroplast leader peptide, in contrast to nuclear NAPs. The chloroplast localization of cNAPL was demonstrated by laser scanning confocal fluorescence microscopy using different chimeric cGFP fusion proteins. Phylogenetic analysis shows that no homologues of cNAPL and its related nuclear counterparts are present in prokaryotic genomes. These data indicate that the chloroplast protein described here is a novel member of the NAP family and most probably has not been acquired from a prokaryotic endosymbiont.

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Transcriptome analysis of the Euglena gracilis plastid chromosome

et al. 2009

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The characterisation of transcript levels of chloroplast genes and their changes under different conditions is an initial step towards understanding chloroplast gene expression and the functional integration of the plastid chromosome into the entire integrated compartmentalised genome of the plant cell. Using RNA from cells of 12 different developmental stages and stress treatments, we have studied the transcript patterns of all 96 genes of the circular plastid chromosome of Euglena gracilis, Pringsheim strain Z, by a macroarray-based approach and Northern analysis of selected genes representing approximately half a dozen operons. The unicellular alga possesses complex, triple-envelope chloroplasts that were acquired by secondary endosymbiosis. (1) Transcripts were detected from all genes, although stationary concentrations varied substantially between individual loci. No obvious economy in the expression pattern with respect to transcription units and genes for complex structures was noted. (2) The chromosome appears to be constitutively expressed under all chosen conditions including stresses such as UV light, temperature, antiplastidial agents, herbicide and heavy metal exposure. (3) The euglenoid organelle transcriptome is qualitatively relatively insensitive to the environment, but exhibited marked overall quantitative changes. The more or less global changes demonstrate that primarily RNA turnover, translational, proteolytic and/or metabolic control regulate organelle gene expression in the alga.

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DNA macroarray and real-time PCR analysis of two nuclear photosystem I mutants from Chlamydomonas reinhardtii reveal downregulation of Lhcb genes but different regulation of Lhca genes

Cited by 5 publications

References 38 publications

Coordinated Regulation of Gene Expression for Carotenoid Metabolism in Chlamydomonas reinhardtii

Coordinated Regulation of Gene Expression for Carotenoid Metabolism in Chlamydomonas reinhardtii

A nucleosome assembly protein-like polypeptide binds to chloroplast group II intron RNA in Chlamydomonas reinhardtii

Transcriptome analysis of the Euglena gracilis plastid chromosome

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