Project description:Identification of target transcripts for the putative chloroplast RNA binding protein CFM2 in Zea mays. CFM2 was immunoprecipitated from a chloroplast extract. Chloroplast extracts were prepared from WT tissue. RNA from the pellet and from the supernatant for each pulldown was labelled with different fluoro-dyes and hybridized onto an array covering the complete maize chloroplast genome. Messages enriched in the immunoprecipitate from WT tissue are likely targets for CFM2.
Project description:Identification of target transcripts for the putative chloroplast RNA binding protein CRP1 in Zea mays. CRP1 was immunoprecipitated from a chloroplast extract. Chloroplast extracts were prepared from WT and CRP1-deficient tissue. RNA from the pellet and from the supernatant for each pulldown was labelled with different fluoro-dyes and hybridized onto an array covering the complete maize chloroplast genome. Messages enriched in the immunoprecipitate from WT tissue, but not enriched in mutant tissue are likely targets for CRP1.
Project description:Chloroplasts contain a dedicated genome, which encodes subunits of the photosynthesis machinery. Transcription of photosynthesis genes is predominantly carried out by a plastid-encoded RNA polymerase (PEP), a nearly 1 MDa complex composed of core subunits with homology to eubacterial RNA polymerases (RNAPs) and at least 12 additional chloroplast-specific PEP-associated proteins (PAPs). However, the architecture of this complex and the functions of the PAPs remain unknown. In this work, a 19-subunit PEP complex from Sinapis alba was purified by glycerol gradient centrifugation. All known subunits of the complex could be identified by LC-MS and the structure of the complex was determined by cryo-electron microscopy.