The S-Phase Cyclin Clb5 Promotes rRNA Gene (rDNA) Stability by Maintaining Replication Initiation Efficiency in rDNA

• Published in: Molecular and cellular biology Vol. 41; no. 5
• Main Authors: Goto, Mayuko, Sasaki, Mariko, Kobayashi, Takehiko
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 22.04.2021; American Society for Microbiology
• Subjects: cell biology; Clb5; Cyclin B - genetics; cyclins; DNA; DNA damage; DNA Damage - genetics; DNA Replication - genetics; DNA, Fungal - genetics; DNA, Ribosomal - genetics; DNA-Binding Proteins - metabolism; genes; Genes, rRNA; genetic instability; genome instability; Genomic Instability - genetics; replication fork arrest; replication origin; Replication Origin - genetics; Research Article; ribosomal RNA gene; S Phase - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; yeasts; replication fork arrest; genome instability; Clb5; replication origin; ribosomal RNA gene
• ISSN: 1098-5549; 0270-7306; 1098-5549
• DOI: 10.1128/MCB.00324-20

Regulation of replication origins is important for complete duplication of the genome, but the effect of origin activation on the cellular response to replication stress is poorly understood. The budding yeast rRNA gene (rDNA) forms tandem repeats and undergoes replication fork arrest at the replication fork barrier (RFB), inducing DNA double-strand breaks (DSBs) and genome instability accompanied by copy number alterations. Here, we demonstrate that the S-phase cyclin Clb5 promotes rDNA stability. Absence of Clb5 led to reduced efficiency of replication initiation in rDNA but had little effect on the number of replication forks arrested at the RFB, suggesting that arrival of the converging fork is delayed and forks are more stably arrested at the RFB. Deletion of CLB5 affected neither DSB formation nor its repair at the RFB but led to homologous recombination-dependent rDNA instability. Therefore, arrested forks at the RFB may be subject to DSB-independent, recombination-dependent rDNA instability. The rDNA instability in clb5Δ was not completely suppressed by the absence of Fob1, which is responsible for fork arrest at the RFB. Thus, Clb5 establishes the proper interval for active replication origins and shortens the travel distance for DNA polymerases, which may reduce Fob1-independent DNA damage.