Cdc14 and Chromosome Condensation: Evaluation of the Recruitment of Condensin to Genomic Regions.
Chromosome condensation is an essential morphological event required for successful DNA segregation during mitosis. The high level of genome compaction achieved during this process is attained by the evolutionary conserved condensin complex. Recently, several lines of evidences have demonstrated that the mitotic phosphatase Cdc14 is required to ensure condensin loading onto chromosomes. To date several approaches have been used in order to characterize condensin activity and regulation, however these techniques are time-consuming and require complex equipment. In this chapter we described an easy and reliable protocol to analyze Cdc14-dependent condensin loading onto specific genomic DNA regions by using a chromatin immunoprecipitation (ChIP) technique.
Cell Cycle and Genome Stability Group, Instituto de Biología Funcional y Genómica, Consejo Superior de Investigaciones Científicas (CSIC), Universidad de Salamanca (USAL), C/ Zacarías González 2, Salamanca, 37007, Spain.
Facundo
F
Ramos
Cell Cycle Group, Medical Research Council, Clinical Sciences Centre, Imperial College London, Du Cane Road, London, W12 0NN, UK.
Joanne
J
Leonard
Cell Cycle and Genome Stability Group, Instituto de Biología Funcional y Genómica, Consejo Superior de Investigaciones Científicas (CSIC), Universidad de Salamanca (USAL), C/ Zacarías González 2, Salamanca, 37007, Spain.
Andrés
A
Clemente-Blanco
Cell Cycle Group, Medical Research Council, Clinical Sciences Centre, Imperial College London, Du Cane Road, London, W12 0NN, UK. luis.aragon@csc.mrc.ac.uk.
Luis
L
Aragón
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D042003:DNA Packaging; D004268:DNA-Binding Proteins / Q000378:metabolism*; D016681:Genome, Fungal; D008938:Mitosis; D046912:Multiprotein Complexes / Q000378:metabolism*; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
C074860:CDC14 protein, S cerevisiae; D018797:Cell Cycle Proteins; D002843:Chromatin; D004268:DNA-Binding Proteins; D046912:Multiprotein Complexes; D029701:Saccharomyces cerevisiae Proteins; C108312:condensin complexes; D017027:Protein Tyrosine Phosphatases; D000251:Adenosine Triphosphatases
Cdc14 phosphatase; Chromatin immunoprecipitation; Chromosome condensation; Condensin; Mitosis