Characterization of a plant mitochondrial active chromosome.
A method is presented for the partial purification of a plant mitochondrial active chromosome (MAC). This method is based on the presence of the mitochondrial chromosome in the insoluble mitochondrial fraction which allows for its rapid purification from the bulk of detergent-solubilized proteins by ultra-centrifugation. The resuspended MAC carrying DNA and RNA-binding proteins retains DNA synthesis and transcription activities comparable to the ones found in isolated mitochondria. In comparison, tRNA-nucleotidyl terminal transferase taken as an example of RNA modifying activities remains in the soluble fraction. MAC purification is proposed as a rapid and efficient first step in the purification of DNA-binding proteins involved in DNA replication and transcription.
Fey
J
J
Institut de Biologie Moléculaire des Plantes du CNRS, UPRA0406, Université Louis Pasteur, Strasbourg, France.
Vermel
M
M
Grienenberger
J
J
Maréchal-Drouard
L
L
Gualberto
J M
JM
D002458:Cell Fractionation; D002875:Chromosomes / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D008928:Mitochondria / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D017393:RNA Editing; D018749:RNA, Plant / Q000302:isolation & purification / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011198:Solanum tuberosum / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D014158:Transcription, Genetic; D014461:Ultracentrifugation; D003313:Zea mays / Q000737:chemistry / Q000201:enzymology / Q000235:genetics
D018749:RNA, Plant; D016601:RNA-Binding Proteins