Search for: All records

Abstract The evolutionarily conserved microspherule protein 1 (MCRS1) has diverse functions, ranging from transcriptional regulation to stabilization of microtubule minus ends in acentrosomal spindles in mammals. A previous study suggested that in the model plant Arabidopsis thaliana, inactivation of an MCRS1 homolog gene led to aborted embryogenesis. To test whether this lethality was caused solely by sporophytic defects, we used the heterozygous emb1967-1/mcrs1-1 mutant for reciprocal crosses with the wild-type plant and found that the MCRS1 gene was dispensable for the development of both male and female gametophytes. An MCRS1–GFP fusion protein was expressed in the mcrs1 mutant and suppressed the mutation as evidenced by restored growth. This functional fusion protein exclusively localized to interphase nuclei and became unnoticeable during mitosis before reappearing in the reforming daughter nuclei. Affinity purification of the MCRS1–GFP protein specifically recovered the Myb-like transcription factor DRMY1 (Development Regulated Myb-like 1) but not microtubule-associated factors. Direct MCRS1–DRMY1 interaction was also demonstrated by a localization-based assay in living cells. Thus, we hypothesized that MCRS1’s function was perhaps linked to transcription factors like DRMY1 and its paralog DP1 for regulation of gene expression during sporophyte development. 

Abstract Spindle assembly in vertebrates requires the Aurora kinase, which is targeted to microtubules and activated by TPX2 (Targeting Protein of XKLP2). In Arabidopsis (Arabidopsis thaliana), TPX2-LIKE 3 (TPXL3), but not the highly conserved TPX2, is essential. To test the hypothesis that TPXL3 regulates the function of α Aurora kinase in spindle assembly, we generated transgenic Arabidopsis lines expressing an artificial microRNA targeting TPXL3 mRNA (amiR-TPXL3). The resulting mutants exhibited growth retardation, which was linked to compromised TPXL3 expression. In the mutant cells, α Aurora was delocalized from spindle microtubules to the cytoplasm, and spindles were assembled without recognizable poles. A functional TPXL3-GFP fusion protein first prominently appeared on the prophase nuclear envelope. Then, TPXL3-GFP localized to spindle microtubules (primarily toward the spindle poles, like γ-tubulin), and finally to the re-forming nuclear envelope during telophase and cytokinesis. However, TPXL3 was absent from phragmoplast microtubules. In addition, we found that the TPXL3 N-terminal Aurora-binding motif, microtubule-binding domain, and importin-binding motif, but not the C-terminal segment, were required for its mitotic function. Expression of truncated TPXL3 variants enhanced the defects in spindle assembly and seedling growth of amiR-TPXL3 plants. Taken together, our findings uncovered the essential function of TPXL3, but not TPX2, in targeting and activating α Aurora kinase for spindle apparatus assembly in Arabidopsis. 

ABSTRACT Plant cytokinesis results in the formation of the cell plate by the phragmoplast which contains dynamic microtubules serving as the track for the delivery of cell wall builders included in Golgi vesicles. During the centrifugal process of cell plate assembly, new microtubules are assembled and bundled at the leading edge to prepare for vesicle transport while older microtubules are disassembled at the lagging edge upon the completion of vesicle delivery. The turnover of phragmoplast microtubules in this process is thought to be regulated by phosphorylation of the key microtubule bundling factor MAP65. A recent study revealed a surprising role of theα‐Aurora kinase, which is typically known for its role in governing the formation of the bipolar spindle apparatus, in phosphorylating the primary microtubule bundler MAP65‐3 in Arabidopsis. This phosphorylation positively contributes to the expansion of the phragmoplast. The phragmoplast midzone is also the hub for other cytokinesis‐important kinases. It is intriguing how these kinases are targeted and how they may crosstalk with each other to orchestrate the expansion of the phragmoplast.