title: Nucleoid-enriched proteomes in developing plastids and chloroplasts from maize leaves: a new conceptual framework for nucleoid functions.
abstract: Plastids contain multiple copies of the plastid chromosome, folded together with proteins and RNA into nucleoids. The degree to which components of the plastid gene expression and protein biogenesis machineries are nucleoid associated, and the factors involved in plastid DNA organization, repair, and replication, are poorly understood. To provide a conceptual framework for nucleoid function, we characterized the proteomes of highly enriched nucleoid fractions of proplastids and mature chloroplasts isolated from the maize (Zea mays) leaf base and tip, respectively, using mass spectrometry. Quantitative comparisons with proteomes of unfractionated proplastids and chloroplasts facilitated the determination of nucleoid-enriched proteins. This nucleoid-enriched proteome included proteins involved in DNA replication, organization, and repair as well as transcription, mRNA processing, splicing, and editing. Many proteins of unknown function, including pentatricopeptide repeat (PPR), tetratricopeptide repeat (TPR), DnaJ, and mitochondrial transcription factor (mTERF) domain proteins, were identified. Strikingly, 70S ribosome and ribosome assembly factors were strongly overrepresented in nucleoid fractions, but protein chaperones were not. Our analysis strongly suggests that mRNA processing, splicing, and editing, as well as ribosome assembly, take place in association with the nucleoid, suggesting that these processes occur cotranscriptionally. The plastid developmental state did not dramatically change the nucleoid-enriched proteome but did quantitatively shift the predominating function from RNA metabolism in undeveloped plastids to translation and homeostasis in chloroplasts. This study extends the known maize plastid proteome by hundreds of proteins, including more than 40 PPR and mTERF domain proteins, and provides a resource for targeted studies on plastid gene expression. Details of protein identification and annotation are provided in the Plant Proteome Database.
authors: Department of Plant Biology, Cornell University, Ithaca, New York 14853, USA.
authors: Wojciech
authors: W
authors: Majeran
authors: Giulia
authors: G
authors: Friso
authors: Yukari
authors: Y
authors: Asakura
authors: Xian
authors: X
authors: Qu
authors: Mingshu
authors: M
authors: Huang
authors: Lalit
authors: L
authors: Ponnala
authors: Kenneth P
authors: KP
authors: Watkins
authors: Alice
authors: A
authors: Barkan
authors: Klaas J
authors: KJ
authors: van Wijk
mesh_terms: D002736:Chloroplasts / Q000378:metabolism*; D004260:DNA Repair / Q000502:physiology; D004261:DNA Replication; D018506:Gene Expression Regulation, Plant; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism; D015388:Organelles / Q000235:genetics / Q000378:metabolism*; D018515:Plant Leaves / Q000254:growth & development* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D018087:Plastids / Q000235:genetics* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017393:RNA Editing; D012326:RNA Splicing; D012260:Ribonucleases / Q000737:chemistry; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism
chemical_list: D024101:Mitochondrial Proteins; D010940:Plant Proteins; D020543:Proteome; D012260:Ribonucleases