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Fast and global reorganization of the chloroplast protein biogenesis network during heat acclimation

ABSTRACT: Heat waves caused by global warming severely challenge photosynthesis and hence plant biomass production, but the mechanisms of its adjustment, repair and biogenesis during heat acclimation are elusive. We provide a global and time-resolved insight, how these processes are modulated during heat acclimation in chloroplasts, revealing profound modulation of protein biogenesis. Ribosome profiling with Chlamydomonas reinhardtii showed early translational downregulation during synthesis of photosynthesis core antenna proteins, a response that is conserved in Nicotiana tabacum. Our data suggest that these early effects may be caused by reduced translocation of ribosomes synthesizing thylakoid proteins, and that later heat acclimation alleviates these effects. Analysis of the proteinaceous interactome of plastid ribosomes demonstrates widespread remodeling of ribosome-associated processes that could be responsible for the early translational impacts and subsequent acclimation responses. The role of chlorophyll metabolism, protein targeting, and assembly of photosystem complexes as co-translational bottlenecks during heat acclimation are discussed.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Chlamydomonas Reinhardtii

TISSUE(S): Cell Culture

SUBMITTER: Markus Räschle

LAB HEAD: Zuzana Storchova

PROVIDER: PXD024949 | Pride | 2022-01-06

REPOSITORIES: Pride

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			Action	DRS
	20190517_HF1_MaRa_SA_P0032_C0012_L5_000_1_01.raw	Raw
	20190517_HF1_MaRa_SA_P0032_C0012_L5_000_1_02.raw	Raw
	20190517_HF1_MaRa_SA_P0032_C0012_L5_000_1_03.raw	Raw
	20190517_HF1_MaRa_SA_P0032_C0012_L5_000_2_01.raw	Raw
	20190517_HF1_MaRa_SA_P0032_C0012_L5_000_2_02.raw	Raw

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The proteomic inventory reveals the chloroplast ribosome as nexus within a diverse protein network

Project description:In plant cells, chloroplast gene expression is predominantly controlled through regulation of translation. Fine-tuning of protein synthesis is vital for plant development, acclimation to environmental challenges and for the assembly of major protein complexes such as the photosynthesis machinery. However, little is known about the degree and nature of the regulatory network, mainly due to challenges associated with the specific isolation of chloroplast ribosomes under conditions that also retain weak or transient interactions. Here, we established a ribosome affinity purification method, which enabled us to map the global interaction network of chloroplast ribosomes. Endogenously tagging a protein of the large or small subunit revealed not only interactors of the holo complex, but also preferential interactors of the two subunits. This includes known canonical regulatory proteins as well as several new proteins belonging to the categories of protein and RNA regulation, photosystem biogenesis, redox control and plastidic metabolism. By further characterizing identified proteins, we provide the first evidence for the existence of a co-translational N-acetyltransferase process in chloroplasts. Our dataset establishes the chloroplast ribosome as nexus in a highly choreographed, spatially interconnected protein network and reveals its wide-ranging regulatory potential during gene expression.

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