Kinesin-5 Mediated Chromosome Congression in Insect Spindles

Emily Tubman, Yungui He, Thomas S. Hays, David J. Odde

Research output: Contribution to journalArticle

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Abstract

Introduction: The microtubule motor protein kinesin-5 is well known to establish the bipolar spindle by outward sliding of antiparallel interpolar microtubules. In yeast, kinesin-5 also facilitates chromosome alignment “congression” at the spindle equator by preferentially depolymerizing long kinetochore microtubules (kMTs). The motor protein kinesin-8 has also been linked to chromosome congression. Therefore, we sought to determine whether kinesin-5 or kinesin-8 facilitates chromosome congression in insect spindles. Methods: RNAi of the kinesin-5 Klp61F and kinesin-8 Klp67A were performed separately in Drosophila melanogaster S2 cells to test for inhibited chromosome congression. Klp61F RNAi, Klp67A RNAi, and control metaphase mitotic spindles expressing fluorescent tubulin and fluorescent Cid were imaged, and their fluorescence distributions were compared. Results: RNAi of Klp61F with a weak Klp61F knockdown resulted in longer kMTs and less congressed kinetochores compared to control over a range of conditions, consistent with kinesin-5 length-dependent depolymerase activity. RNAi of the kinesin-8 Klp67A revealed that kMTs relative to the spindle lengths were not longer compared to control, but rather that the spindles were longer, indicating that Klp67A acts preferentially as a length-dependent depolymerase on interpolar microtubules without significantly affecting kMT length and chromosome congression. Conclusions: This study demonstrates that in addition to establishing the bipolar spindle, kinesin-5 regulates kMT length to facilitate chromosome congression in insect spindles. It expands on previous yeast studies, and it expands the role of kinesin-5 to include kMT assembly regulation in eukaryotic mitosis.

Original languageEnglish (US)
Pages (from-to)25-36
Number of pages12
JournalCellular and Molecular Bioengineering
Volume11
Issue number1
DOIs
StatePublished - Feb 1 2018

Fingerprint

Kinesin
Chromosomes, Human, Pair 5
Microtubules
Chromosomes
Chromosome
Insects
Kinetochores
RNA Interference
Yeast
Expand
Proteins
Protein
Equator
Antiparallel
Dependent
Drosophilidae
Fluorescence
Yeasts
Alignment
Microtubule Proteins

Keywords

  • Kinesin-5
  • Microtubules
  • Mitosis

PubMed: MeSH publication types

  • Journal Article

Cite this

Kinesin-5 Mediated Chromosome Congression in Insect Spindles. / Tubman, Emily; He, Yungui; Hays, Thomas S.; Odde, David J.

In: Cellular and Molecular Bioengineering, Vol. 11, No. 1, 01.02.2018, p. 25-36.

Research output: Contribution to journalArticle

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