Description

• Title:
  Ribosomal DNA status inferred from DNA cloud assays and mass spectrometry identification of agarose-squeezed proteins interacting with chromatin (ASPIC-MS)
• Creator:
  Krol, Kamil;
  Jendrysek, Justyna;
  Dębski, Janusz;
  Skoneczny, Marek;
  Kurlandzka, Anna;
  Kaminska, Joanna;
  Dadlez, Michal;
  Skoneczna, Adrianna
• Description:
  Ribosomal RNA-encoding genes (rDNA) are the most abundant genes in eukaryotic genomes. To meet the high demand for rRNA, rDNA genes are present in multiple tandem repeats clustered on a single or several chromosomes and are vastly transcribed. To facilitate intensive transcription and prevent rDNA destabilization, the rDNA-encoding portion of the chromosome is confined in the nucleolus. However, the rDNA region is susceptible to recombination and DNA damage, accumulating mutations, rearrangements and atypical DNA structures. Various sophisticated techniques have been applied to detect these abnormalities. Here, we present a simple method for the evaluation of the activity and integrity of an rDNA region called a “DNA cloud assay”. We verified the efficacy of this method using yeast mutants lacking genes important for nucleolus function and maintenance (RAD52, SGS1, RRM3, PIF1, FOB1 and RPA12). The DNA cloud assay permits the evaluation of nucleolus status and is compatible with downstream analyses, such as the chromosome comet assay to identify DNA structures present in the cloud and mass spectrometry of agarose squeezed proteins (ASPIC-MS) to detect nucleolar DNA-bound proteins, including Las17, the homolog of human Wiskott-Aldrich Syndrome Protein (WASP)
• Object availability:
  free access
• Rights:
  cc_by
• Date:
  2017.04.11
• Type:
  Article;
  PeerReviewed
• Language:
  English
• Format:
  application/pdf
• Publisher:
  Impact Journals
• Relation: http://eprints.ibb.waw.pl/1334 ;http://www.impactjournals.com/oncotarget ;
  10.18632/oncotarget.15332
• Subject:
  QH301 Biology;
  QH426 Genetics
• Identifier: http://eprints.ibb.waw.pl/1334/1/oncotarget-08-24988.pdf ;
  Krol, Kamil and Jendrysek, Justyna and Dębski, Janusz and Skoneczny, Marek and Kurlandzka, Anna and Kaminska, Joanna and Dadlez, Michal and Skoneczna, Adrianna (2017) Ribosomal DNA status inferred from DNA cloud assays and mass spectrometry identification of agarose-squeezed proteins interacting with chromatin (ASPIC-MS). Oncotarget, 8 (15). pp. 24988-25004. ISSN 1949-2553
• Data provider:
  Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk
• Can I use it?:
  check license
• Type:
  other