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Kinesin-4 KIF21B is a potent microtubule pausing factor

View Article: PubMed Central - PubMed

ABSTRACT

Microtubules are dynamic polymers that in cells can grow, shrink or pause, but the factors that promote pausing are poorly understood. Here, we show that the mammalian kinesin-4 KIF21B is a processive motor that can accumulate at microtubule plus ends and induce pausing. A few KIF21B molecules are sufficient to induce strong growth inhibition of a microtubule plus end in vitro. This property depends on non-motor microtubule-binding domains located in the stalk region and the C-terminal WD40 domain. The WD40-containing KIF21B tail displays preference for a GTP-type over a GDP-type microtubule lattice and contributes to the interaction of KIF21B with microtubule plus ends. KIF21B also contains a motor-inhibiting domain that does not fully block the interaction of the protein with microtubules, but rather enhances its pause-inducing activity by preventing KIF21B detachment from microtubule tips. Thus, KIF21B combines microtubule-binding and regulatory activities that together constitute an autonomous microtubule pausing factor.

Doi:: http://dx.doi.org/10.7554/eLife.24746.001

No MeSH data available.


(A) Far-UV CD spectra (inset) and thermal unfolding profile of recombinant KIF21B rCC. CD measurements, performed in PBS at a protein concentration of 0.166 mg/ml. (B) Oligomerization state of recombinant KIF21B rCC determined by sedimentation velocity AUC at 20°C and at three different protein concentrations.DOI:http://dx.doi.org/10.7554/eLife.24746.034
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fig6s4: (A) Far-UV CD spectra (inset) and thermal unfolding profile of recombinant KIF21B rCC. CD measurements, performed in PBS at a protein concentration of 0.166 mg/ml. (B) Oligomerization state of recombinant KIF21B rCC determined by sedimentation velocity AUC at 20°C and at three different protein concentrations.DOI:http://dx.doi.org/10.7554/eLife.24746.034

Mentions: 10.7554/eLife.24746.030Figure 6—figure supplement 4—source data 1.An excel sheet with numerical data on the quantification of far-UV CD spectra (inset) and thermal unfolding profile of recombinant KIF21B rCC1 represented as plots in Figure 6—figure supplement 4A.


Kinesin-4 KIF21B is a potent microtubule pausing factor
(A) Far-UV CD spectra (inset) and thermal unfolding profile of recombinant KIF21B rCC. CD measurements, performed in PBS at a protein concentration of 0.166 mg/ml. (B) Oligomerization state of recombinant KIF21B rCC determined by sedimentation velocity AUC at 20°C and at three different protein concentrations.DOI:http://dx.doi.org/10.7554/eLife.24746.034
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383399&req=5

fig6s4: (A) Far-UV CD spectra (inset) and thermal unfolding profile of recombinant KIF21B rCC. CD measurements, performed in PBS at a protein concentration of 0.166 mg/ml. (B) Oligomerization state of recombinant KIF21B rCC determined by sedimentation velocity AUC at 20°C and at three different protein concentrations.DOI:http://dx.doi.org/10.7554/eLife.24746.034
Mentions: 10.7554/eLife.24746.030Figure 6—figure supplement 4—source data 1.An excel sheet with numerical data on the quantification of far-UV CD spectra (inset) and thermal unfolding profile of recombinant KIF21B rCC1 represented as plots in Figure 6—figure supplement 4A.

View Article: PubMed Central - PubMed

ABSTRACT

Microtubules are dynamic polymers that in cells can grow, shrink or pause, but the factors that promote pausing are poorly understood. Here, we show that the mammalian kinesin-4 KIF21B is a processive motor that can accumulate at microtubule plus ends and induce pausing. A few KIF21B molecules are sufficient to induce strong growth inhibition of a microtubule plus end in vitro. This property depends on non-motor microtubule-binding domains located in the stalk region and the C-terminal WD40 domain. The WD40-containing KIF21B tail displays preference for a GTP-type over a GDP-type microtubule lattice and contributes to the interaction of KIF21B with microtubule plus ends. KIF21B also contains a motor-inhibiting domain that does not fully block the interaction of the protein with microtubules, but rather enhances its pause-inducing activity by preventing KIF21B detachment from microtubule tips. Thus, KIF21B combines microtubule-binding and regulatory activities that together constitute an autonomous microtubule pausing factor.

Doi:: http://dx.doi.org/10.7554/eLife.24746.001

No MeSH data available.