Title | Claps | Level | Year | L/Y |
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Systems Biology Approach in Chlamydomonas Reveals Connections between Copper Nutrition and Multiple Metabolic Steps[C][W][OA]
12 auth. M. Castruita, D. Casero, Steven J. Karpowicz, J. Kropat, Astrid Vieler, Scott I. Hsieh, ...
RNA-seq assessment of the transcriptome of autotrophic and heterotrophic Chlamydomonas as a function of copper nutrition reveals changes in redox metabolism regulated by CRR1, an SBP domain transcription factor. The changes in RNA abundance impact t…
RNA-seq assessment of the transcriptome of autotrophic and heterotrophic Chlamydomonas as a function of copper nutrition reveals changes in redox metabolism regulated by CRR1, an SBP domain transcription factor. The changes in RNA abundance impact the abundance of specific plastid-localized proteins and the level of saturation of plastid galactolipids. In this work, we query the Chlamydomonas reinhardtii copper regulon at a whole-genome level. Our RNA-Seq data simulation and analysis pipeline validated a 2-fold cutoff and 10 RPKM (reads per kilobase of mappable length per million mapped reads) (~1 mRNA per cell) to reveal 63 CRR1 targets plus another 86 copper-responsive genes. Proteomic and immunoblot analyses captured 25% of the corresponding proteins, whose abundance was also dependent on copper nutrition, validating transcriptional regulation as a major control mechanism for copper signaling in Chlamydomonas. The impact of copper deficiency on the expression of several O2-dependent enzymes included steps in lipid modification pathways. Quantitative lipid profiles indicated increased polyunsaturation of fatty acids on thylakoid membrane digalactosyldiglycerides, indicating a global impact of copper deficiency on the photosynthetic apparatus. Discovery of a putative plastid copper chaperone and a membrane protease in the thylakoid suggest a mechanism for blocking copper utilization in the chloroplast. We also found an example of copper sparing in the N assimilation pathway: the replacement of copper amine oxidase by a flavin-dependent backup enzyme. Forty percent of the targets are previously uncharacterized proteins, indicating considerable potential for new discovery in the biology of copper.
Published in
The Plant Cell
|
3
|
6 | 2011 |
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