CLASPs function redundantly to regulate astral microtubules in the C. elegans embryo

• Published in: Developmental biology 2012-08, Vol.368 (2), p.242-254
• Main Authors: Espiritu, Eugenel B., Krueger, Lori E., Ye, Anna, Rose, Lesilee S.
• Format: Article
• Language: English
• Subjects: Animals; Animals, Genetically Modified; Asymmetric division; binding proteins; Blotting, Western; Caenorhabditis elegans; Caenorhabditis elegans - embryology; Caenorhabditis elegans - genetics; Caenorhabditis elegans - metabolism; Caenorhabditis elegans Proteins - genetics; Caenorhabditis elegans Proteins - metabolism; Caenorhabditis elegans Proteins - physiology; chromosome segregation; cortex; Cytoplasm - metabolism; Embryo, Nonmammalian - cytology; Embryo, Nonmammalian - embryology; Embryo, Nonmammalian - metabolism; embryogenesis; genome; Immunohistochemistry; kinetochores; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; meiosis; Microscopy, Confocal; Microtubule-Associated Proteins - genetics; Microtubule-Associated Proteins - metabolism; Microtubule-Associated Proteins - physiology; Microtubules; Microtubules - metabolism; Mitosis; molecular motor proteins; polymerization; RNA Interference; Spindle Apparatus - metabolism; Spindle positioning; Time Factors
• ISSN: 0012-1606; 1095-564X
• DOI: 10.1016/j.ydbio.2012.05.016

Microtubule dynamics are thought to play an important role in regulating microtubule interactions with cortical force generating motor proteins that position the spindle during asymmetric cell division. CLASPs are microtubule-associated proteins that have a conserved role in regulating microtubule dynamics in diverse cell types. Caenorhabditis elegans has three CLASP homologs in its genome. CLS-2 is known to localize to kinetochores and is needed for chromosome segregation at meiosis and mitosis; however CLS-1 and CLS-3 have not been reported to have any role in embryonic development. Here, we show that depletion of CLS-2 in combination with either CLS-1 or CLS-3 results in defects in nuclear rotation, maintenance of spindle length, and spindle displacement in the one-cell embryo. Polarity is normal in these embryos, but reduced numbers of astral microtubules reach all regions of the cortex at the time of spindle positioning. Analysis of the microtubule plus-end tracker EB1 also revealed a reduced number of growing microtubules reaching the cortex in CLASP depleted embryos, but the polymerization rate of astral microtubules was not slower than in wild type. These results indicate that C. elegans CLASPs act partially redundantly to regulate astral microtubules and position the spindle during asymmetric cell division. Further, we show that these spindle pole-positioning roles are independent of the CLS-2 binding proteins HCP-1 and HCP-2. ► CLASPs function redundantly in C. elegans embryos during spindle positioning. ► CLASP depleted embryos have reduced numbers of microtubule-cortex contacts. ► The CLS-2 binding proteins HCP-1 and 2 are not required for spindle positioning.