Deletion of a trypanosome telomere leads to loss of silencing and progressive loss of terminal DNA in the absence of cell cycle arrest.
Eukaryotic chromosomes are capped with telomeres which allow complete chromosome replication and prevent the ends from being recognized by the repair machinery. The African trypanosome, Trypanosoma brucei, is a protozoan parasite where antigenic variation requires reversible silencing of a repository of telomere-adjacent variant surface glycoprotein (VSG) genes. We have investigated the role of the telomere adjacent to a repressed VSG. In cells lacking telomerase, the rate of telomere-repeat loss appeared to be inversely proportional to telomere length. We therefore constructed strains in which a single telomere could be immediately removed by conditional I-SceI meganuclease cleavage. Following telomere deletion, cells maintain and segregate the damaged chromosome without repairing it. These cells continue to proliferate at the normal rate but progressively lose terminal DNA at the broken end. Although sirtuin-dependent repression is lost along with the telomere, VSG-silencing is preserved. The results provide direct evidence for telomere-dependent repression but suggest a telomere-independent mode of VSG-silencing. They also indicate the absence of a telomere-loss checkpoint in T. brucei.
Glover
Lucy
L
London School of Hygiene and Tropical Medicine, Keppel Street, London, WC1E 7HT, UK.
Alsford
Sam
S
Beattie
Caroline
C
Horn
David
D
D000818:Animals; D002453:Cell Cycle; D002872:Chromosome Deletion; D004260:DNA Repair; D016054:DNA, Protozoan / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D017353:Gene Deletion; D020868:Gene Silencing; D029701:Saccharomyces cerevisiae Proteins; D037761:Sirtuins / Q000378:metabolism; D019098:Telomerase / Q000235:genetics; D016615:Telomere / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D014346:Trypanosoma brucei brucei / Q000235:genetics* / Q000378:metabolism; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000235:genetics / Q000378:metabolism
D016054:DNA, Protozoan; D029701:Saccharomyces cerevisiae Proteins; D014643:Variant Surface Glycoproteins, Trypanosoma; D019098:Telomerase; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific; D037761:Sirtuins