Mitotic chromosome condensation in the rDNA requires TRF4 and DNA topoisomerase I in Saccharomyces cerevisiae.
DNA topoisomerase I (topo I) is known to participate in the process of DNA replication, but is not essential in Saccharomyces cerevisiae. The TRF4 gene is also nonessential and was identified in a screen for mutations that are inviable in combination with a top1 null mutation. Here we report the surprising finding that a top1 trf4-ts double mutant is defective in the mitotic events of chromosome condensation, spindle elongation, and nuclear segregation, but not in DNA replication. Direct examination of rDNA-containing mitotic chromosomes demonstrates that a top1 trf4-ts mutant fails both to establish and to maintain chromosome condensation in the rDNA at mitosis. We show that the Trf4p associates physically with both Smclp and Smc2p, the S. cerevisiae homologs of Xenopus proteins that are required for mitotic chromosome condensation in vitro. The defect in the top1 trf4-ts mutant is sensed by the MAD1-dependent spindle assembly checkpoint but not by the RAD9-dependent DNA damage checkpoint, further supporting the notion that chromosome structure influences spindle assembly. These data indicate that TOP1 (encoding topo I) and TRF4 participate in overlapping or dependent steps in mitotic chromosome condensation and serve to define a previously unrecognized biological function of topo I.
Department of Radiation Oncology, University of California, San Francisco 94143, USA.
I B
IB
Castaño
P M
PM
Brzoska
B U
BU
Sadoff
H
H
Chen
M F
MF
Christman
D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus; D002503:Centromere; D002868:Chromosomal Proteins, Non-Histone; D002875:Chromosomes; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000378:metabolism*; D004275:DNA, Ribosomal; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D008938:Mitosis; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D015398:Signal Transduction; D008941:Spindle Apparatus / Q000502:physiology; D000098550:Structural Maintenance of Chromosome Protein 1
D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004275:DNA, Ribosomal; D005656:Fungal Proteins; D009687:Nuclear Proteins; C090455:SMC2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D000098550:Structural Maintenance of Chromosome Protein 1; D004264:DNA Topoisomerases, Type I