Newly Characterized Region of CP190 Associates with Microtubules and Mediates Proper Spindle Morphology in Drosophila Stem Cells

Plevock, Karen M.; Galletta, Brian J.; Slep, Kevin C.; Rusan, Nasser M.; Prigent, Claude

doi:10.1371/journal.pone.0144174

Newly Characterized Region of CP190 Associates with Microtubules and Mediates Proper Spindle Morphology in Drosophila Stem Cells

Journal Article · Wed Dec 09 00:00:00 EST 2015 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0144174· OSTI ID:1245866

Plevock, Karen M. ^[1]; Galletta, Brian J. ^[2]; Slep, Kevin C. ^[3]; Rusan, Nasser M. ^[2]; Prigent, Claude ^[4]

National Inst. of Health, Bethesda, MD (United States); Univ. of North Carolina, Chapel Hill, NC (United States)
National Inst. of Health, Bethesda, MD (United States)
Univ. of North Carolina, Chapel Hill, NC (United States)
Inst. de Génétique et Développement de Rennes (France)

CP190 is a large, multi-domain protein, first identified as a centrosome protein with oscillatory localization over the course of the cell cycle. During interphase it has a well-established role within the nucleus as a chromatin insulator. Upon nuclear envelope breakdown, there is a striking redistribution of CP190 to centrosomes and the mitotic spindle, in addition to the population at chromosomes. Here, we investigate CP190 in detail by performing domain analysis in cultured Drosophila S2 cells combined with protein structure determination by X-ray crystallography, in vitro biochemical characterization, and in vivo fixed and live imaging of cp190 mutant flies. Our analysis of CP190 identifies a novel N-terminal centrosome and microtubule (MT) targeting region, sufficient for spindle localization. This region consists of a highly conserved BTB domain and a linker region that serves as the MT binding domain. We present the 2.5 Å resolution structure of the CP190 N-terminal 126 amino acids, which adopts a canonical BTB domain fold and exists as a stable dimer in solution. The ability of the linker region to robustly localize to MTs requires BTB domain-mediated dimerization. Deletion of the linker region using CRISPR significantly alters spindle morphology and leads to DNA segregation errors in the developing Drosophila brain neuroblasts. Collectively, we highlight a multivalent MT-binding architecture in CP190, which confers distinct subcellular cytoskeletal localization and function during mitosis.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: National Heart Lung and Blood Inst.; National Inst. of Health

OSTI ID:: 1245866

Journal Information:: PLoS ONE, Journal Name: PLoS ONE Journal Issue: 12 Vol. 10; ISSN 1932-6203

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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59 BASIC BIOLOGICAL SCIENCES
Centrosomes
Crystal structure
Dimerization
Dimers (Chemical physics)
Drosophila melanogaster
Insulators
MTS assay
Mitosis

Newly Characterized Region of CP190 Associates with Microtubules and Mediates Proper Spindle Morphology in Drosophila Stem Cells

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Cited By (6)

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