Biology subjects

Goshima, G.

Publications and source records attributed to Goshima, G..

5 recordsLinked to original sources

bioRxiv2018

Chromosome missegregation causes somatic polyploidization in plants

Lagging chromosome is a hallmark of aneuploidy arising from errors in the kinetochore-spindle attachment in animal cells. However, kinetochore components and cellular phenotypes associated with kinetochore dysfunction are much less explored in plants. Here, we carried out a comprehensive characterization of conserved kinetochore components in the moss Physcomitrella patens and uncovered a distinct scenario in plant cells regarding both the localization and cellular impact of the kinetochore proteins. Most surprisingly, knock-down of several kinetochore proteins led to polyploidy, not aneuploidy, through cytokinesis failure in >90% of the cells that exhibited lagging chromosomes for several minutes or longer. The resultant cells, containing two or more nuclei, proceeded to the next cell cycle and eventually developed into polyploid plants. As lagging chromosomes have been observed in various plant species in the wild, our observation raised a possibility that they could be one of the natural pathways to polyploidy in plants.

cell biology↗

bioRxiv2018

Drosophila kinesin-8 stabilises kinetochore-microtubule interaction

Kinesin-8 is required for proper chromosome alignment in a variety of animal and yeast cell types. However, how this conserved motor protein controls chromosome alignment remains unclear, as multiple biochemical activities, including inconsistent ones between studies, have been identified for this motor family. Here, we show that Drosophila kinesin-8 Klp67A possesses both microtubule (MT) plus-end-stabilising and -destabilising activities in addition to commonly observed MT plus-end-directed motility and tubulin-binding activity in vitro, and is required for stable kinetochore-MT attachment during prometaphase in S2 cells In the absence of kinesin-8Klp67A, abnormally-long MTs interact in an \"end-on\" fashion with kinetochores at normal frequency. However, the interaction was not stable and, once-attached, MTs were frequently detached. This phenotype was rescued by ectopic expression of MT plus-end-stabilising factor CLASP, but not by artificial shortening of MTs. These results suggest that MT-stabilising activity of kinesin-8Klp67A is critical for stable kinetochore-MT attachment. Finally, human kinesin-8KIF18A was also shown important to ensure proper MT attachment.

cell biology↗

bioRxiv2018

KCH kinesin drives nuclear transport and cytoskeletal coalescence for tip cell growth

Long-distance transport along microtubules (MTs) is critical for intracellular organisation. In animals, antagonistic motor proteins kinesin (plus end-directed) and dynein (minus end-directed) drive cargo transport. In land plants, however, the identity of motors responsible for transport is poorly understood, as genes encoding cytoplasmic dynein are missing. How other functions of dynein are brought about in plants also remains unknown. Here, we show that a subclass of the kinesin-14 family, KCH--which can also bind actin--drives MT minus end-directed nuclear transport in the moss Physcomitrella patens. When all four KCH genes were deleted, the nucleus was not maintained in the cell centre but was translocated to the apical end of protonemal cells. In the knockout (KO) line, apical cell tip growth was also severely suppressed. KCH was localised on MTs, including at the MT focal point near the tip where MT plus ends coalesced with actin filaments. MT focus was not persistent in KCH KO lines, whereas actin destabilisation also disrupted the focus despite KCH remaining on unfocused MTs. Functions of nuclear transport and tip growth were distinct, as a truncated KCH construct restored nuclear transport activity but not tip growth retardation of the KO line. Thus, our study identified KCH as a long-distance retrograde transporter as well as a cytoskeletal crosslinker, reminiscent of the versatile animal dynein.

cell biology↗

bioRxiv2017

Multiple kinesin-14 family members drive microtubule minus-end-directed transport in plant cells

Minus-end-directed cargo transport along microtubules (MTs) is exclusively driven by the molecular motor dynein in a wide variety of cell types. Interestingly, plants have lost the genes encoding dynein during evolution; the MT motors that compensate for dynein function are unknown. Here, we show that two members of the kinesin-14 family drive minus-end-directed transport in plants. Gene knockout analyses of the moss Physcomitrella patens revealed that the plant-specific class-VI kinesin-14, KCBP, is required for minus-end-directed transport of the nucleus and chloroplasts. Purified KCBP directly bound to acidic phospholipids (PLs) and unidirectionally transported PL liposomes along MTs in vitro. Thus, minus-end-directed transport of membranous cargoes might be driven by their direct interaction with this motor protein. Newly nucleated cytoplasmic MTs represent another known cargo exhibiting minus-end-directed motility, and we identified the conserved class-I kinesin-14 (ATK) as the motor involved. These results suggest that kinesin-14 motors were duplicated and developed as alternative MT-based minus-end-directed transporters in land plants.

cell biology↗

bioRxiv2017

Human microcephaly ASPM protein is a spindle pole-focusing factor

Nonsense mutations in the ASPM gene have been most frequently identified among familial microcephaly patients. Depletion of the Drosophila orthologue causes spindle pole unfocusing during mitosis in multiple cell types. However, it remains unknown whether human ASPM has a similar function. Here, using CRISPR-based gene knockout (KO) and RNA interference combined with auxin-inducible degron, we show that ASPM functions in spindle pole organisation during mitotic metaphase redundantly with another microcephaly protein CDK5RAP2 (also called CEP215) in human tissue culture cells. Deletion of the ASPM gene alone did not affect spindle morphology or mitotic progression. However, when the pericentriolar material protein CDK5RAP2 was depleted in ASPM KO cells, spindle poles were unfocused during prometaphase and anaphase onset was significantly delayed. The phenotypic analysis of CDK5RAP2-depleted cells suggested that the pole-focusing function of CDK5RAP2 is independent of its known function to localise the kinesin-14 motor HSET or activate the{gamma} -tubulin complex. Finally, a hypomorphic mutation identified in ASPM microcephaly patients similarly caused spindle pole unfocusing in the absence of CDK5RAP2, suggesting a possible link between spindle pole disorganisation and microcephaly.

cell biology↗