Project description:Here we show that the phytochrome-less chlorophyte Chlamydomonas reinhardtii retains a functional pathway to synthesize the linear tetrapyrrole (bilin) precursor of the phytochrome chromophore. Reverse genetic, metabolic inactivation and bilin rescue experiments establish that this pathway is needed for heme iron acquisition and for the diurnal transition to phototrophic growth. RNA-Seq measurements reveal a bilin-dependent signaling network that is necessary for the heterotrophic to phototrophic transition. These results imply the presence of a novel bilin sensor pathway that may be widely distributed amongst oxygenic photosynthetic organisms.
Project description:RNA populations in Chlamydomonas reinhardtii Keywords: Highly parallel pyrosequencing Small RNAs were prepared from Chlamydomonas reinhardtii total extracts,ligated to a 3' adaptor and a 5' acceptor sequentially, and then RT-PCR amplified. PCR products were reamplified using a pair of 454 cloning primers and provided to 454 Life Sciences (Branford, CT) for sequencing. For technical details, see Tao Zhao, Guanglin Li, Shijun Mi, Shan Li, Gregory J. Hannon, Xiu-Jie Wang, and Yijun Qi. 2007. A Complex System of Small RNAs in the Unicellular Green Alga Chlamydomonas reinhardtii. Genes & Development
Project description:Here we show that the phytochrome-less chlorophyte Chlamydomonas reinhardtii retains a functional pathway to synthesize the linear tetrapyrrole (bilin) precursor of the phytochrome chromophore. Reverse genetic, metabolic inactivation and bilin rescue experiments establish that this pathway is needed for heme iron acquisition and for the diurnal transition to phototrophic growth. RNA-Seq measurements reveal a bilin-dependent signaling network that is necessary for the heterotrophic to phototrophic transition. These results imply the presence of a novel bilin sensor pathway that may be widely distributed amongst oxygenic photosynthetic organisms. We isolated RNA from heterotrophic suspension cultures of 4A+ WT and the hmox1 mutant grown in the presence or absence of 0.1 mM BV IXM-NM-1 before and after transfer to low light.
Project description:Here, we report a transcriptomics analysis on a day in the life of Chlamydomonas reinhardtii. Cultures of this unicellular alga were grown in photobioreactors on a 12 h light / 12 h dark cycle. Samples were collected at regular intervals and subjected to a transcriptomics analysis by RNA-Seq.
Project description:endogenous small RNAs from Chlamydomonas reinhardtii strain J3(mt-) vegetative cells Keywords: High throughput 454 small RNA sequencing
Project description:The tetrapyrroles Mg-protoporphyrin IX (MgProto) and heme, in Chlamydomonas reinhardtii only synthesized within the chloroplast, have been implicated in retrograde control of nuclear gene expression in this unicellular green alga. However, feeding of the two tetrapyrroles to Chlamydomonas cultures growing in the dark has previously been shown to transiently induce 5 nuclear genes, among them three genes coding for the chloroplast heat shock proteins HSP70A, B and E. Here, we measured the impact of MgProto and hemin feeding in the dark on changes in gene expression at the genomic level. About 10% of the 10000 genes represented on the microarray showed a transient up or down regulation with a fold change of 4 or more (p ≤0.05). The two most prominent groups of regulated genes were those where both MgProto and heme caused either an up or down regulation. Minor regulatory groups consisted of genes which were either down- or up-regulated by one of the tetrapyrroles but not by the other. In contrast, feeding of protoporphyrin IX had no regulatory effect on a number of selected genes. Interestingly, 499 of the 982 responding genes were also regulated by heat shock; 85% of those showed the same response (up or down) as seen after MgProto/heme feeding, indicating a previously not anticipated role of MgProto and heme in stress response. Indeed, most prominent among the functional groups of annotated genes up or down regulated by the tetrapyrroles were those whose gene products are involved in protein folding and/or protein degradation. Striking is the virtual absence of regulated genes that encode constituents of the photosynthetic apparatus. This and the transient nature of changes in gene expression observed upon feeding of the tetrapyrroles suggest a signaling role of these plastid compounds in the adaptation of the alga to alterations in the environment.