Katanin maintains meiotic metaphase chromosome alignment and spindle structure in vivo and has multiple effects on microtubules in vitro

Karen McNally, Evan Berg, Daniel B. Cortes, Veronica Hernandez, Paul E. Mains, Francis J. McNally

Research output: Contribution to journalArticle

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Abstract

Assembly of Caenorhabditis elegans female meiotic spindles requires both MEI-1 and MEI-2 subunits of the microtubule-severing ATPase katanin. Strong loss-of-function mutants assemble apolar intersecting microtubule arrays, whereas weaker mutants assemble bipolar meiotic spindles that are longer than wild type. To determine whether katanin is also required for spindle maintenance, we monitored metaphase I spindles after a fast-acting mei-1(ts) mutant was shifted to a nonpermissive temperature. Within 4 min of temperature shift, bivalents moved off the metaphase plate, and microtubule bundles within the spindle lengthened and developed a high degree of curvature. Spindles eventually lost bipolar structure. Immunofluorescence of embryos fixed at increasing temperature indicated that MEI-1 was lost from spindle microtubules before loss of ASPM-1, indicating that MEI-1 and ASPM-1 act independently at spindle poles. We quantified the microtubule-severing activity of purified MEI-1/MEI-2 complexes corresponding to six different point mutations and found a linear relationship between microtubule disassembly rate and meiotic spindle length. Previous work showed that katanin is required for severing at points where two microtubules intersect in vivo. We show that purified MEI-1/MEI-2 complexes preferentially sever at intersections between two microtubules and directly bundle microtubules in vitro. These activities could promote parallel/antiparallel microtubule organization in meiotic spindles.

Original languageEnglish (US)
Pages (from-to)1037-1049
Number of pages13
JournalMolecular Biology of the Cell
Volume25
Issue number7
DOIs
StatePublished - Apr 1 2014

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Metaphase
Microtubules
Chromosomes
Spindle Apparatus
Temperature
katanin
In Vitro Techniques
Spindle Poles
Caenorhabditis elegans
Point Mutation
Fluorescent Antibody Technique
Adenosine Triphosphatases
Embryonic Structures
Maintenance

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Katanin maintains meiotic metaphase chromosome alignment and spindle structure in vivo and has multiple effects on microtubules in vitro. / McNally, Karen; Berg, Evan; Cortes, Daniel B.; Hernandez, Veronica; Mains, Paul E.; McNally, Francis J.

In: Molecular Biology of the Cell, Vol. 25, No. 7, 01.04.2014, p. 1037-1049.

Research output: Contribution to journalArticle

McNally, Karen ; Berg, Evan ; Cortes, Daniel B. ; Hernandez, Veronica ; Mains, Paul E. ; McNally, Francis J. / Katanin maintains meiotic metaphase chromosome alignment and spindle structure in vivo and has multiple effects on microtubules in vitro. In: Molecular Biology of the Cell. 2014 ; Vol. 25, No. 7. pp. 1037-1049.
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