title: Telomerase-associated protein 1, HSP90, and topoisomerase IIalpha associate directly with the BLM helicase in immortalized cells using ALT and modulate its helicase activity using telomeric DNA substrates.
abstract: The BLM helicase associates with the telomere structural proteins TRF1 and TRF2 in immortalized cells using the alternative lengthening of telomere (ALT) pathways. This work focuses on identifying protein partners of BLM in cells using ALT. Mass spectrometry and immunoprecipitation techniques have identified three proteins that bind directly to BLM and TRF2 in ALT cells: telomerase-associated protein 1 (TEP1), heat shock protein 90 (HSP90), and topoisomerase IIalpha (TOPOIIalpha). BLM predominantly co-localizes with these proteins in foci actively synthesizing DNA during late S and G(2)/M phases of the cell cycle when ALT is thought to occur. Immunoprecipitation studies also indicate that only HSP90 and TOPOIIalpha are components of a specific complex containing BLM, TRF1, and TRF2 but that this complex does not include TEP1. TEP1, TOPOIIalpha, and HSP90 interact directly with BLM in vitro and modulate its helicase activity on telomere-like DNA substrates but not on non-telomeric substrates. Initial studies suggest that knockdown of BLM in ALT cells reduces average telomere length but does not do so in cells using telomerase.
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University College of Medicine, Columbus, Ohio 43210-2207, USA.
authors.forename: Saumitri
authors.initials: S
authors.lastname: Bhattacharyya
authors.forename: Jeremy
authors.initials: J
authors.lastname: Keirsey
authors.forename: Beatriz
authors.initials: B
authors.lastname: Russell
authors.forename: Juraj
authors.initials: J
authors.lastname: Kavecansky
authors.forename: Kate
authors.initials: K
authors.lastname: Lillard-Wetherell
authors.forename: Kambiz
authors.initials: K
authors.lastname: Tahmaseb
authors.forename: John J
authors.initials: JJ
authors.lastname: Turchi
authors.forename: Joanna
authors.initials: J
authors.lastname: Groden