Dataset Information

Separation and Paired Proteome Profiling of Plant Chloroplast and Cytoplasmic Ribosomes

ABSTRACT: Conventional preparation methods of plant ribosomes fail to resolve non-translating chloroplast or cytoplasmic ribosome subunits from translating fractions. We established preparation of these ribosome complexes from Arabidopsis thaliana leaf, root, and seed tissues by optimized sucrose density gradient centrifugation of protease protected plant extracts. The method co-purified non-translating 30S and 40S ribosome subunits, separated non-translating 50S from 60S subunits, and resolved assembled monosomes from low oligomeric polysomes. Combining ribosome fractionation with microfluidic rRNA analysis and proteomics, we characterized the rRNA and ribosomal protein (RP) composition. The identity of cytoplasmic and chloroplast ribosome complexes and the presence of ribosome biogenesis factors in the 60S - 80S sedimentation interval were verified. In vivo cross-linking of leaf tissue stabilized ribosome biogenesis complexes but induced polysome run-off. Omitting cross-linking, the established paired fractionation and proteome analysis monitored relative abundances of plant chloroplast and cytoplasmic ribosome fractions and enabled analysis of RP composition and ribosome associated proteins including transiently associated biogenesis factors.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Rosette

SUBMITTER: Michal Gorka

LAB HEAD: Joachim Kopka

PROVIDER: PXD019329 | Pride | 2021-09-09

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	08_02_18_REIL1_REIL2_File1.raw	Raw
	08_02_18_REIL1_REIL2_File2.raw	Raw
	08_02_18_REIL1_REIL2_File3.raw	Raw
	08_02_18_REIL1_REIL2_File4.raw	Raw
	08_02_18_REIL1_REIL2_File5.raw	Raw

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Publications

Separation and Paired Proteome Profiling of Plant Chloroplast and Cytoplasmic Ribosomes.

Firmino Alexandre Augusto Pereira AAP Gorka Michal M Graf Alexander A Skirycz Aleksandra A Martinez-Seidel Federico F Zander Kerstin K Kopka Joachim J Beine-Golovchuk Olga O

Plants (Basel, Switzerland) 20200714 7

Conventional preparation methods of plant ribosomes fail to resolve non-translating chloroplast or cytoplasmic ribosome subunits from translating fractions. We established preparation of these ribosome complexes from <i>Arabidopsis thaliana</i> leaf, root, and seed tissues by optimized sucrose density gradient centrifugation of protease protected plant extracts. The method co-purified non-translating 30S and 40S ribosome subunits separated non-translating 50S from 60S subunits, and resolved asse ...[more]

PMID: 32674508

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