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Kinetochore-dependent microtubule rescue ensures their efficient and sustained interactions in early mitosis.

Gandhi SR, Gierliński M, Mino A, Tanaka K, Kitamura E, Clayton L, Tanaka TU - Dev. Cell (2011)

Bottom Line: Meanwhile, microtubule rescue distal to the kinetochore is also promoted by Stu2, which is transported by a kinesin-8 motor Kip3 along the microtubule from the kinetochore.Microtubule extension following rescue facilitates interaction with other widely scattered kinetochores, diminishing long delays in collecting the complete set of kinetochores by microtubules.Thus, kinetochore-dependent microtubule rescue ensures efficient and sustained kinetochore-microtubule interactions in early mitosis.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Centre for Gene Regulation & Expression, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.

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Microtubule Rescue Distal to the Kinetochore Is Promoted by Stu2 Transport from the Kinetochore along the Microtubule Lateral Surface to Its Plus End(A) MT rescue distal to the KT is distinct from the growth of an overlapping MT, and is preceded by Stu2 transport from the KT to the MT end. Time-lapse images of PMET3-CDC20 PGAL-CEN3-tetOs TetR-GFP YFP-TUB1 STU2-3×CFP (T4986) cells showing (i) Stu2 transport leading to MT rescue distal to the KT, and (ii) the growth of an overlapping MT. Schematic diagrams are shown at the bottom of the images. The graph shows MT length (orange), CEN3–spindle-pole distance (blue), and the position of Stu2 during its transport (i, magenta) and Stu2 at the end of an overlapping MT (ii, green). The cells were treated in the same way as in Figure 1C. Images were collected every 5 s using a common channel for GFP and YFP, and a separate channel for CFP. Zero time is set arbitrarily for the first panel. Scale bar, 1 μm.(B) Transport of a hypomorphic stu2 mutant from the KT to the MT end often fails to promote MT rescue distal to the KT. (i) Time-lapse images, in which Stu2-ΔTOG1 transport did not lead to MT rescue. PMET3-CDC20 PGAL-CEN3-tetOs TetR-3×CFP CFP-TUB1 STU2+ PSTU2-stu2-ΔTOG1-3×GFP (T9242) cells were treated as in Figure 1C, and images were collected every 7 s for CFP and GFP. Zero time is set arbitrarily for the first panel. Scale bar, 1 μm. (ii) The graph is plotted as in (Ai). (iii) The table shows the number of events in which Stu2 transport to the end of a shrinking MT did or did not lead to MT rescue distal to CEN3.See also Figure S2.
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fig3: Microtubule Rescue Distal to the Kinetochore Is Promoted by Stu2 Transport from the Kinetochore along the Microtubule Lateral Surface to Its Plus End(A) MT rescue distal to the KT is distinct from the growth of an overlapping MT, and is preceded by Stu2 transport from the KT to the MT end. Time-lapse images of PMET3-CDC20 PGAL-CEN3-tetOs TetR-GFP YFP-TUB1 STU2-3×CFP (T4986) cells showing (i) Stu2 transport leading to MT rescue distal to the KT, and (ii) the growth of an overlapping MT. Schematic diagrams are shown at the bottom of the images. The graph shows MT length (orange), CEN3–spindle-pole distance (blue), and the position of Stu2 during its transport (i, magenta) and Stu2 at the end of an overlapping MT (ii, green). The cells were treated in the same way as in Figure 1C. Images were collected every 5 s using a common channel for GFP and YFP, and a separate channel for CFP. Zero time is set arbitrarily for the first panel. Scale bar, 1 μm.(B) Transport of a hypomorphic stu2 mutant from the KT to the MT end often fails to promote MT rescue distal to the KT. (i) Time-lapse images, in which Stu2-ΔTOG1 transport did not lead to MT rescue. PMET3-CDC20 PGAL-CEN3-tetOs TetR-3×CFP CFP-TUB1 STU2+ PSTU2-stu2-ΔTOG1-3×GFP (T9242) cells were treated as in Figure 1C, and images were collected every 7 s for CFP and GFP. Zero time is set arbitrarily for the first panel. Scale bar, 1 μm. (ii) The graph is plotted as in (Ai). (iii) The table shows the number of events in which Stu2 transport to the end of a shrinking MT did or did not lead to MT rescue distal to CEN3.See also Figure S2.

Mentions: Is MT rescue distal to the KT also dependent on Stu2, similar to rescue that occurs at the KT? Indeed, as we reported previously, MT rescue distal to CEN3 always happened coincidentally (24 out of 24 events) with arrival of Stu2 at the MT plus end after it had been transported from CEN3 along the MT lateral surface (Tanaka et al., 2005) (Figure 3Ai). Stu2 transport along a MT was not a misinterpretation of the Stu2 signal at the end of an overlapping, growing MT, which could be discerned by a partly thicker MT signal (Figure 3Aii, 75–165 s) and a prior Stu2 movement from a spindle pole to CEN3 (Figure 3Aii, 75–90 s). This conclusion was also confirmed by a separate experiment, using photobleaching of a MT region (Figure S2A). It appeared that Stu2 transport from a KT occurred and led to MT rescue distal to the KT not only in the engineered assay system (Figure 3Ai), but also in physiological conditions (Figure S2B). Stu2 transport from CEN3 along a MT was observed with similar frequency during MT growth and shrinkage in the engineered assay (data not shown). If Stu2 arrived at the end of a growing MT, there was no obvious effect.


Kinetochore-dependent microtubule rescue ensures their efficient and sustained interactions in early mitosis.

Gandhi SR, Gierliński M, Mino A, Tanaka K, Kitamura E, Clayton L, Tanaka TU - Dev. Cell (2011)

Microtubule Rescue Distal to the Kinetochore Is Promoted by Stu2 Transport from the Kinetochore along the Microtubule Lateral Surface to Its Plus End(A) MT rescue distal to the KT is distinct from the growth of an overlapping MT, and is preceded by Stu2 transport from the KT to the MT end. Time-lapse images of PMET3-CDC20 PGAL-CEN3-tetOs TetR-GFP YFP-TUB1 STU2-3×CFP (T4986) cells showing (i) Stu2 transport leading to MT rescue distal to the KT, and (ii) the growth of an overlapping MT. Schematic diagrams are shown at the bottom of the images. The graph shows MT length (orange), CEN3–spindle-pole distance (blue), and the position of Stu2 during its transport (i, magenta) and Stu2 at the end of an overlapping MT (ii, green). The cells were treated in the same way as in Figure 1C. Images were collected every 5 s using a common channel for GFP and YFP, and a separate channel for CFP. Zero time is set arbitrarily for the first panel. Scale bar, 1 μm.(B) Transport of a hypomorphic stu2 mutant from the KT to the MT end often fails to promote MT rescue distal to the KT. (i) Time-lapse images, in which Stu2-ΔTOG1 transport did not lead to MT rescue. PMET3-CDC20 PGAL-CEN3-tetOs TetR-3×CFP CFP-TUB1 STU2+ PSTU2-stu2-ΔTOG1-3×GFP (T9242) cells were treated as in Figure 1C, and images were collected every 7 s for CFP and GFP. Zero time is set arbitrarily for the first panel. Scale bar, 1 μm. (ii) The graph is plotted as in (Ai). (iii) The table shows the number of events in which Stu2 transport to the end of a shrinking MT did or did not lead to MT rescue distal to CEN3.See also Figure S2.
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fig3: Microtubule Rescue Distal to the Kinetochore Is Promoted by Stu2 Transport from the Kinetochore along the Microtubule Lateral Surface to Its Plus End(A) MT rescue distal to the KT is distinct from the growth of an overlapping MT, and is preceded by Stu2 transport from the KT to the MT end. Time-lapse images of PMET3-CDC20 PGAL-CEN3-tetOs TetR-GFP YFP-TUB1 STU2-3×CFP (T4986) cells showing (i) Stu2 transport leading to MT rescue distal to the KT, and (ii) the growth of an overlapping MT. Schematic diagrams are shown at the bottom of the images. The graph shows MT length (orange), CEN3–spindle-pole distance (blue), and the position of Stu2 during its transport (i, magenta) and Stu2 at the end of an overlapping MT (ii, green). The cells were treated in the same way as in Figure 1C. Images were collected every 5 s using a common channel for GFP and YFP, and a separate channel for CFP. Zero time is set arbitrarily for the first panel. Scale bar, 1 μm.(B) Transport of a hypomorphic stu2 mutant from the KT to the MT end often fails to promote MT rescue distal to the KT. (i) Time-lapse images, in which Stu2-ΔTOG1 transport did not lead to MT rescue. PMET3-CDC20 PGAL-CEN3-tetOs TetR-3×CFP CFP-TUB1 STU2+ PSTU2-stu2-ΔTOG1-3×GFP (T9242) cells were treated as in Figure 1C, and images were collected every 7 s for CFP and GFP. Zero time is set arbitrarily for the first panel. Scale bar, 1 μm. (ii) The graph is plotted as in (Ai). (iii) The table shows the number of events in which Stu2 transport to the end of a shrinking MT did or did not lead to MT rescue distal to CEN3.See also Figure S2.
Mentions: Is MT rescue distal to the KT also dependent on Stu2, similar to rescue that occurs at the KT? Indeed, as we reported previously, MT rescue distal to CEN3 always happened coincidentally (24 out of 24 events) with arrival of Stu2 at the MT plus end after it had been transported from CEN3 along the MT lateral surface (Tanaka et al., 2005) (Figure 3Ai). Stu2 transport along a MT was not a misinterpretation of the Stu2 signal at the end of an overlapping, growing MT, which could be discerned by a partly thicker MT signal (Figure 3Aii, 75–165 s) and a prior Stu2 movement from a spindle pole to CEN3 (Figure 3Aii, 75–90 s). This conclusion was also confirmed by a separate experiment, using photobleaching of a MT region (Figure S2A). It appeared that Stu2 transport from a KT occurred and led to MT rescue distal to the KT not only in the engineered assay system (Figure 3Ai), but also in physiological conditions (Figure S2B). Stu2 transport from CEN3 along a MT was observed with similar frequency during MT growth and shrinkage in the engineered assay (data not shown). If Stu2 arrived at the end of a growing MT, there was no obvious effect.

Bottom Line: Meanwhile, microtubule rescue distal to the kinetochore is also promoted by Stu2, which is transported by a kinesin-8 motor Kip3 along the microtubule from the kinetochore.Microtubule extension following rescue facilitates interaction with other widely scattered kinetochores, diminishing long delays in collecting the complete set of kinetochores by microtubules.Thus, kinetochore-dependent microtubule rescue ensures efficient and sustained kinetochore-microtubule interactions in early mitosis.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Centre for Gene Regulation & Expression, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.

Show MeSH
Related in: MedlinePlus