The γ-tubulin-specific inhibitor gatastatin reveals temporal requirements of microtubule nucleation during the cell cycle.
Inhibitors of microtubule (MT) assembly or dynamics that target α/β-tubulin are widely exploited in cancer therapy and biological research. However, specific inhibitors of the MT nucleator γ-tubulin that would allow testing temporal functions of γ-tubulin during the cell cycle are yet to be identified. By evolving β-tubulin-binding drugs we now find that the glaziovianin A derivative gatastatin is a γ-tubulin-specific inhibitor. Gatastatin decreased interphase MT dynamics of human cells without affecting MT number. Gatastatin inhibited assembly of the mitotic spindle in prometaphase. Addition of gatastatin to preformed metaphase spindles altered MT dynamics, reduced the number of growing MTs and shortened spindle length. Furthermore, gatastatin prolonged anaphase duration by affecting anaphase spindle structure, indicating the continuous requirement of MT nucleation during mitosis. Thus, gatastatin facilitates the dissection of the role of γ-tubulin during the cell cycle and reveals the sustained role of γ-tubulin.
Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8572, Japan.|Zentrum für Molekulare Biologie der Universität Heidelberg, ZMBH-DKFZ Alliance, Im Neuenheimer Feld 282, Heidelberg 69120, Germany.
Takumi
T
Chinen
Zentrum für Molekulare Biologie der Universität Heidelberg, ZMBH-DKFZ Alliance, Im Neuenheimer Feld 282, Heidelberg 69120, Germany.
Peng
P
Liu
Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8571, Japan.
Shuya
S
Shioda
Zentrum für Molekulare Biologie der Universität Heidelberg, ZMBH-DKFZ Alliance, Im Neuenheimer Feld 282, Heidelberg 69120, Germany.
Judith
J
Pagel
Zentrum für Molekulare Biologie der Universität Heidelberg, ZMBH-DKFZ Alliance, Im Neuenheimer Feld 282, Heidelberg 69120, Germany.
Berati
B
Cerikan
Zentrum für Molekulare Biologie der Universität Heidelberg, ZMBH-DKFZ Alliance, Im Neuenheimer Feld 282, Heidelberg 69120, Germany.
Tien-Chen
TC
Lin
Zentrum für Molekulare Biologie der Universität Heidelberg, ZMBH-DKFZ Alliance, Im Neuenheimer Feld 282, Heidelberg 69120, Germany.
Oliver
O
Gruss
Department of Medicinal Chemistry, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392, Japan.
Yoshiki
Y
Hayashi
Department of Medicinal Chemistry, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392, Japan.
Haruka
H
Takeno
Laboratory for Cell Polarity Regulation, RIKEN Quantitative Biology Center, Suita 565-0874, Japan.
Tomohiro
T
Shima
Laboratory for Cell Polarity Regulation, RIKEN Quantitative Biology Center, Suita 565-0874, Japan.
Yasushi
Y
Okada
Division of Applied Chemistry, Graduate School of Natural Science and Technology, Okayama University, 3-1-1 Tsushima-naka, Kita-ku, Okayama 700-8530, Japan.
Ichiro
I
Hayakawa
Department of Medicinal Chemistry, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392, Japan.
Yoshio
Y
Hayashi
Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8571, Japan.
Hideo
H
Kigoshi
Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8572, Japan.
Takeo
http://orcid.org/0000-0003-2377-7499
T
Usui
Zentrum für Molekulare Biologie der Universität Heidelberg, ZMBH-DKFZ Alliance, Im Neuenheimer Feld 282, Heidelberg 69120, Germany.
Elmar
E
Schiebel
D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D006801:Humans; D007529:Isoflavones / Q000737:chemistry / Q000494:pharmacology*; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism*; D008941:Spindle Apparatus / Q000187:drug effects / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism; D050257:Tubulin Modulators / Q000737:chemistry / Q000494:pharmacology*
D007529:Isoflavones; D014404:Tubulin; D050257:Tubulin Modulators; C521560:glaziovianin A