title: Human CST abundance determines recovery from diverse forms of DNA damage and replication stress.
abstract: Mammalian CST (CTC1-STN1-TEN1) is a telomere-associated complex that functions in telomere duplex replication and fill-in synthesis of the telomeric C-strand following telomerase action. CST also facilitates genome-wide replication recovery after HU-induced fork stalling by increasing origin firing. CTC1 and STN1 were originally isolated as a DNA polymerase α stimulatory factor. Here we explore how CST abundance affects recovery from drugs that cause different types of DNA damage and replication stress. We show that recovery from HU and aphidicolin induced replication stress is increased by CST over-expression. Elevated CST increases dNTP incorporation and origin firing after HU release and decreases the incidence of anaphase bridges and micronuclei after aphidicolin removal. While the frequency of origin firing after HU release is proportional to CST abundance, the number of cells entering S-phase to initiate replication is unchanged by CST overexpression or STN1 depletion. Instead the CST-related changes in origin firing take place in cells that were already in S-phase at the time of HU addition, indicating that CST modulates firing of late or dormant origins. CST abundance also influences cell viability after treatment with HU, aphidicolin, MMS and camptothecin. Viability is increased by elevated CST and decreased by STN1 depletion, indicating that endogenous CST levels are limiting. However, CST abundance does not affect viability after MMC treatment. Thus, CST facilitates recovery from many, but not all, forms of exogenous DNA damage. Overall our results suggest that CST is needed in stoichiometric amounts to facilitate re-initiation of DNA replication at repaired forks and/or dormant origins.
authors.affiliation: a Department of Cancer Biology ; University of Cincinnati ; Cincinnati , OH USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.forename: Jason
authors.initials: J
authors.lastname: Stewart
authors.forename: Carolyn M
authors.initials: CM
authors.lastname: Price
mesh_terms: D000818:Animals; D002135:Calcium-Binding Proteins / Q000235:genetics*; D004249:DNA Damage / Q000187:drug effects* / Q000235:genetics; D004256:DNA Polymerase I; D004261:DNA Replication / Q000187:drug effects* / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D006918:Hydroxyurea / Q000633:toxicity; D019098:Telomerase / Q000235:genetics; D016615:Telomere / Q000235:genetics; D034501:Telomere-Binding Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: C503225:CLSTN1 protein, human; D002135:Calcium-Binding Proteins; C544946:Stn1 protein, human; D034501:Telomere-Binding Proteins; C544947:Ten1 protein, human; D019098:Telomerase; D004256:DNA Polymerase I; D006918:Hydroxyurea
keywords: CPT, camptothecin; CST, CTC1-STN1-TEN1; CTC1; CldU, chlorodeoxyuridine; DNA repair; DNA replication; EdU, ethenyl deoxyuridine; HU, hydroxyurea; IdU, iododeoxyuridine; MMC, mitomycin C; MMS, methyl methanesulfonate; MTS, multiple telomere signals; STN1; TEN1; replication origin; telomere