BuGZ is required for Bub3 stability, Bub1 kinetochore function, and chromosome alignment.
During mitosis, the spindle assembly checkpoint (SAC) monitors the attachment of kinetochores (KTs) to the plus ends of spindle microtubules (MTs) and prevents anaphase onset until chromosomes are aligned and KTs are under proper tension. Here, we identify a SAC component, BuGZ/ZNF207, from an RNAi viability screen in human glioblastoma multiforme (GBM) brain tumor stem cells. BuGZ binds to and stabilizes Bub3 during interphase and mitosis through a highly conserved GLE2p-binding sequence (GLEBS) domain. Inhibition of BuGZ results in loss of both Bub3 and its binding partner Bub1 from KTs, reduction of Bub1-dependent phosphorylation of centromeric histone H2A, attenuation of KT-based Aurora B kinase activity, and lethal chromosome congression defects in cancer cells. Phylogenetic analysis indicates that BuGZ orthologs are highly conserved among eukaryotes, but are conspicuously absent from budding and fission yeasts. These findings suggest that BuGZ has evolved to facilitate Bub3 activity and chromosome congression in higher eukaryotes.
Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98195, USA.
Chad M
CM
Toledo
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523, USA.
Jacob A
JA
Herman
Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada.
Jonathan B
JB
Olsen
Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Yu
Y
Ding
Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Philip
P
Corrin
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Emily J
EJ
Girard
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
James M
JM
Olson
Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada.
Andrew
A
Emili
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523, USA. Electronic address: jennifer.deluca@colostate.edu.
Jennifer G
JG
DeLuca
Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98195, USA. Electronic address: paddison@fhcrc.org.
Patrick J
PJ
Paddison
D000595:Amino Acid Sequence; D000818:Animals; D064107:Aurora Kinase B / Q000378:metabolism; D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002877:Chromosomes, Human / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D005909:Glioblastoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D006657:Histones / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D018386:Kinetochores / Q000378:metabolism*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis / Q000502:physiology; D008969:Molecular Sequence Data; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology; D010766:Phosphorylation; D010802:Phylogeny; D000075223:Poly-ADP-Ribose Binding Proteins; D055550:Protein Stability; D034741:RNA, Small Interfering / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D008941:Spindle Apparatus / Q000502:physiology*; D016335:Zinc Fingers / Q000235:genetics*
C489444:BUB3 protein, human; D018797:Cell Cycle Proteins; D006657:Histones; D008869:Microtubule-Associated Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D034741:RNA, Small Interfering; C576921:AURKB protein, human; D064107:Aurora Kinase B