Telomere length kinetics assay (TELKA) sorts the telomere length maintenance (tlm) mutants into functional groups
2014 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 42, no 10, 6314-6325 p.Article in journal (Refereed) Published
Genome-wide systematic screens in yeast have uncovered a large gene network (the telomere length maintenance network or TLM), encompassing more than 400 genes, which acts coordinatively to maintain telomere length. Identifying the genes was an important first stage; the next challenge is to decipher their mechanism of action and to organize then into functional groups or pathways. Here we present a new telomere-length measuring program, TelQuant, and a novel assay, telomere length kinetics assay, and use them to organize tlm mutants into functional classes. Our results show that a mutant defective for the relatively unknown MET7 gene has the same telomeric kinetics as mutants defective for the ribonucleotide reductase subunit Rnr1, in charge of the limiting step in dNTP synthesis, or for the Ku heterodimer, a well-established telomere complex. We confirm the epistatic relationship between the mutants and show that physical interactions exist between Rnr1 and Met7. We also show that Met7 and the Ku heterodimer affect dNTP formation, and play a role in non-homologous end joining. Thus, our telomere kinetics assay uncovers new functional groups, as well as complex genetic interactions between tlm mutants.
Place, publisher, year, edition, pages
Oxford University Press, 2014. Vol. 42, no 10, 6314-6325 p.
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject Molecular Medicine
IdentifiersURN: urn:nbn:se:umu:diva-87935DOI: 10.1093/nar/gku267ISI: 000338768100024OAI: oai:DiVA.org:umu-87935DiVA: diva2:712386
FunderKnut and Alice Wallenberg FoundationSwedish Cancer Society