Stefan Schmollinger1, Timo Mühlhaus2, Nanette R Boyle1, Ian K Blaby1, David Casero3, Tabea Mettler4, Jeffrey L Moseley5, Janette Kropat1, Frederik Sommer2, Daniela Strenkert1, Dorothea Hemme2, Matteo Pellegrini6, Arthur R Grossman5, Mark Stitt4, Michael Schroda2, Sabeeha S Merchant7. 1. Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095. 2. Molecular Biotechnology and Systems Biology, Technische Universität Kaiserslautern, D-67663 Kaiserslautern, Germany Max Planck Institute of Molecular Plant Physiology, D-14476 Potsdam-Golm, Germany. 3. Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, California 90095. 4. Max Planck Institute of Molecular Plant Physiology, D-14476 Potsdam-Golm, Germany. 5. Department of Plant Biology, Carnegie Institution for Science, Stanford, California 94305. 6. Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, California 90095 Institute of Genomics and Proteomics, University of California, Los Angeles, California 90095. 7. Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095 Institute of Genomics and Proteomics, University of California, Los Angeles, California 90095 merchant@chem.ucla.edu.
Abstract
Nitrogen (N) is a key nutrient that limits global primary productivity; hence, N-use efficiency is of compelling interest in agriculture and aquaculture. We used Chlamydomonas reinhardtii as a reference organism for a multicomponent analysis of the N starvation response. In the presence of acetate, respiratory metabolism is prioritized over photosynthesis; consequently, the N-sparing response targets proteins, pigments, and RNAs involved in photosynthesis and chloroplast function over those involved in respiration. Transcripts and proteins of the Calvin-Benson cycle are reduced in N-deficient cells, resulting in the accumulation of cycle metabolic intermediates. Both cytosolic and chloroplast ribosomes are reduced, but via different mechanisms, reflected by rapid changes in abundance of RNAs encoding chloroplast ribosomal proteins but not cytosolic ones. RNAs encoding transporters and enzymes for metabolizing alternative N sources increase in abundance, as is appropriate for the soil environmental niche of C. reinhardtii. Comparison of the N-replete versus N-deplete proteome indicated that abundant proteins with a high N content are reduced in N-starved cells, while the proteins that are increased have lower than average N contents. This sparing mechanism contributes to a lower cellular N/C ratio and suggests an approach for engineering increased N-use efficiency.
Nitrogen (N) is a key nutrient that limits global primary productivity; hence, N-use efficiency is of compelling interest in agriculture and aquaculture. We used n class="Species">Chlamydomonas reinhardtii as a reference organism for a multicomponent analysis of the N starvation response. In the presence of acetate, respiratory metabolism is prioritized over photosynthesis; consequently, the N-sparing response targets proteins, pigments, and RNAs involved in photosynthesis and chloroplast function over those involved in respiration. Transcripts and proteins of the Calvin-Benson cycle are reduced in N-deficient cells, resulting in the accumulation of cycle metabolic intermediates. Both cytosolic and chloroplast ribosomes are reduced, but via different mechanisms, reflected by rapid changes in abundance of RNAs encoding chloroplast ribosomal proteins but not cytosolic ones. RNAs encoding transporters and enzymes for metabolizing alternative N sources increase in abundance, as is appropriate for the soil environmental niche of C. reinhardtii. Comparison of the N-replete versus N-deplete proteome indicated that abundant proteins with a high N content are reduced in N-starved cells, while the proteins that are increased have lower than average N contents. This sparing mechanism contributes to a lower cellular N/C ratio and suggests an approach for engineering increased N-use efficiency.
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