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

Journal Article · · PLoS ONE
 [1];  [2];  [3];  [2];  [4]
  1. National Inst. of Health, Bethesda, MD (United States); Univ. of North Carolina, Chapel Hill, NC (United States)
  2. National Inst. of Health, Bethesda, MD (United States)
  3. Univ. of North Carolina, Chapel Hill, NC (United States)
  4. Inst. de Génétique et Développement de Rennes (France)
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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.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
National Inst. of Health; National Heart Lung and Blood Inst.
Grant/Contract Number:
R01GM094415; T32GM008570; 1ZIAHL006104
OSTI ID:
1245866
Journal Information:
PLoS ONE, Vol. 10, Issue 12; ISSN 1932-6203
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 14 works
Citation information provided by
Web of Science

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Cited By (6)

Opbp is a new architectural/insulator protein required for ribosomal gene expression journal September 2017
HIPP1 stabilizes the interaction between CP190 and Su(Hw) in the Drosophila insulator complex journal December 2019
Role of Su(Hw) zinc finger 10 and interaction with CP190 and Mod(mdg4) proteins in recruiting the Su(Hw) complex to chromatin sites in Drosophila journal February 2018
The same domain of Su(Hw) is required for enhancer blocking and direct promoter repression journal March 2019
Mechanisms of Enhancer-Promoter Interactions in Higher Eukaryotes journal January 2021
The Centrioles, Centrosomes, Basal Bodies, and Cilia of Drosophila melanogaster journal May 2017

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Related Subjects

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