Proteomics

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Characterization vy crosslinking-mass spectrometry of plastid-encoded RNA polymerase purified from Sinapis alba.


ABSTRACT: RNA polymerases (RNAPs) transcribe DNA into RNA and are found in all living organisms with several degrees of complexity from single polypeptide chain to multimeric enzymes. In chloroplast of angiosperms, two RNAPs are involved in plastid gene transcription: the nuclear-encoded RNA polymerase (NEP) and the plastid-encoded RNA polymerase (PEP). The PEP is a prokaryotic-type multimeric RNAP found in different states depending on light stimuli and cell identity. One of these active states requires the assembly of nuclear-encoded proteins named PEP-Associated Proteins (PAPs) with the catalytic core, triggering the timely transcription of photosynthesis-associated plastid genes in cells acquiring their photosynthetic apparatus. A purification procedure was used to enrich native PEP from Sinapis alba chloroplasts. Crosslinking coupled to mass spectrometry provided initial structural information about the relative position of PEP subunits within the complex.

INSTRUMENT(S): Orbitrap Exploris 480, Q Exactive HF

ORGANISM(S): Sinapis Alba

TISSUE(S): Cotyledon

SUBMITTER: Yohann Couté  

LAB HEAD: Yohann Couté

PROVIDER: PXD032739 | Pride | 2022-09-12

REPOSITORIES: Pride

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Publications

Three-Dimensional Envelope and Subunit Interactions of the Plastid-Encoded RNA Polymerase from <i>Sinapis alba</i>.

Ruedas Rémi R   Muthukumar Soumiya Sankari SS   Kieffer-Jaquinod Sylvie S   Gillet François-Xavier FX   Fenel Daphna D   Effantin Grégory G   Pfannschmidt Thomas T   Couté Yohann Y   Blanvillain Robert R   Cobessi David D  

International journal of molecular sciences 20220831 17


RNA polymerases (RNAPs) are found in all living organisms. In the chloroplasts, the plastid-encoded RNA polymerase (PEP) is a prokaryotic-type multimeric RNAP involved in the selective transcription of the plastid genome. One of its active states requires the assembly of nuclear-encoded PEP-Associated Proteins (PAPs) on the catalytic core, producing a complex of more than 900 kDa, regarded as essential for chloroplast biogenesis. In this study, sequence alignments of the catalytic core subunits  ...[more]

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