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Toward a Molecular Mechanism of Bran...

Alfaro-Aco, Raymundo .

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Toward a Molecular Mechanism of Branching Microtubule Nucleation.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Toward a Molecular Mechanism of Branching Microtubule Nucleation./
Author:	Alfaro-Aco, Raymundo .
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2020,
Description:	149 p.
Notes:	Source: Dissertations Abstracts International, Volume: 81-09, Section: B.
Contained By:	Dissertations Abstracts International81-09B.
Subject:	Cellular biology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=27671424
ISBN:	9781392853290

Toward a Molecular Mechanism of Branching Microtubule Nucleation.
Alfaro-Aco, Raymundo .

Toward a Molecular Mechanism of Branching Microtubule Nucleation. - Ann Arbor : ProQuest Dissertations & Theses, 2020 - 149 p.

Source: Dissertations Abstracts International, Volume: 81-09, Section: B.

Thesis (Ph.D.)--Princeton University, 2020.

This item must not be sold to any third party vendors.

The microtubule cytoskeleton supports cell function by giving cells their shape, organizing their interior and segregating chromosomes. Microtubules are nucleated from specific locations at precise times in the cell cycle. However, the factors that constitute these microtubule nucleation pathways and their mode of action still need to be identified. Recently, it was uncovered that microtubules nucleate from pre-existing microtubules within the mitotic spindle, which requires the proteins TPX2, augmin and gamma-TuRC, but the molecular mechanism was unknown. Here, I investigate the role of TPX2 in branching microtubule nucleation, and via an in vitro reconstitution, demonstrate that the three factors TPX2, augmin and gamma-TuRC are sufficient to initiate this process.The C-terminal region of TPX2 is the minimal domain capable of stimulating branching microtubule nucleation in Xenopus egg extract. It contains newly identified gamma-TuRC nucleation activator motifs that are necessary for the function of the protein. While it is dispensable for branching microtubule nucleation, the N-terminal region of TPX2 enhances the efficiency of the reaction. Furthermore, TPX2 has the unexpected ability to directly recruit gamma-TuRC and augmin to pre-existing microtubules. In an in vitro reconstitution, TPX2 and augmin enable gamma-TuRC-dependent microtubule nucleation at preferred branching angles of less than 90 degrees from regularly spaced patches along microtubules. This work elucidates how the microtubule nucleator gamma-TuRC is localized and activated to cause branching microtubule nucleation, a process that is critical for spindle assembly. I anticipate these results will help explain how other microtubule nucleation pathways give rise to specific microtubule architectures in the cell.

ISBN: 9781392853290Subjects--Topical Terms:

3172791
Cellular biology.
Subjects--Index Terms:

Branching

Toward a Molecular Mechanism of Branching Microtubule Nucleation.
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100 1 $a Alfaro-Aco, Raymundo . $3 3547453
245 1 0 $a Toward a Molecular Mechanism of Branching Microtubule Nucleation.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2020
300 $a 149 p.
500 $a Source: Dissertations Abstracts International, Volume: 81-09, Section: B.
500 $a Advisor: Petry, Sabine.
502 $a Thesis (Ph.D.)--Princeton University, 2020.
506 $a This item must not be sold to any third party vendors.
506 $a This item must not be added to any third party search indexes.
520 $a The microtubule cytoskeleton supports cell function by giving cells their shape, organizing their interior and segregating chromosomes. Microtubules are nucleated from specific locations at precise times in the cell cycle. However, the factors that constitute these microtubule nucleation pathways and their mode of action still need to be identified. Recently, it was uncovered that microtubules nucleate from pre-existing microtubules within the mitotic spindle, which requires the proteins TPX2, augmin and gamma-TuRC, but the molecular mechanism was unknown. Here, I investigate the role of TPX2 in branching microtubule nucleation, and via an in vitro reconstitution, demonstrate that the three factors TPX2, augmin and gamma-TuRC are sufficient to initiate this process.The C-terminal region of TPX2 is the minimal domain capable of stimulating branching microtubule nucleation in Xenopus egg extract. It contains newly identified gamma-TuRC nucleation activator motifs that are necessary for the function of the protein. While it is dispensable for branching microtubule nucleation, the N-terminal region of TPX2 enhances the efficiency of the reaction. Furthermore, TPX2 has the unexpected ability to directly recruit gamma-TuRC and augmin to pre-existing microtubules. In an in vitro reconstitution, TPX2 and augmin enable gamma-TuRC-dependent microtubule nucleation at preferred branching angles of less than 90 degrees from regularly spaced patches along microtubules. This work elucidates how the microtubule nucleator gamma-TuRC is localized and activated to cause branching microtubule nucleation, a process that is critical for spindle assembly. I anticipate these results will help explain how other microtubule nucleation pathways give rise to specific microtubule architectures in the cell.
590 $a School code: 0181.
650 4 $a Cellular biology. $3 3172791
650 4 $a Biochemistry. $3 518028
653 $a Branching
653 $a Cytoskeleton
653 $a Microtubule
653 $a Nucleation
653 $a Reconstitution
653 $a Spindle
690 $a 0379
690 $a 0487
710 2 $a Princeton University. $b Molecular Biology. $3 2101701
773 0 $t Dissertations Abstracts International $g 81-09B.
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791 $a Ph.D.
792 $a 2020
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=27671424