Document Detail


An inducible RNA interference system in Physcomitrella patens reveals a dominant role of augmin in phragmoplast microtubule generation.
MedLine Citation:
PMID:  22505727     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Mitosis is a fundamental process of eukaryotic cell proliferation. However, the molecular mechanisms underlying mitosis remain poorly understood in plants partly because of the lack of an appropriate model cell system in which loss-of-function analyses can be easily combined with high-resolution microscopy. Here, we developed an inducible RNA interference (RNAi) system and three-dimensional time-lapse confocal microscopy in the moss Physcomitrella patens that allowed in-depth phenotype characterization of the moss genes essential for cell division. We applied this technique to two microtubule regulators, augmin and γ-tubulin complexes, whose mitotic roles remain obscure in plant cells. Live imaging of caulonemal cells showed that they proceed through mitosis with continual generation and self-organization of acentrosomal microtubules. We demonstrated that augmin plays an important role in γ-tubulin localization and microtubule generation from prometaphase to cytokinesis. Most evidently, microtubule formation in phragmoplasts was severely compromised after RNAi knockdown of an augmin subunit, leading to incomplete expansion of phragmoplasts and cytokinesis failure. Knockdown of the γ-tubulin complex affected microtubule formation throughout mitosis. We conclude that postanaphase microtubule generation is predominantly stimulated by the augmin/γ-tubulin machinery in moss and further propose that this RNAi system serves as a powerful tool to dissect the molecular mechanisms underlying mitosis in land plants.
Authors:
Yuki Nakaoka; Tomohiro Miki; Ryuta Fujioka; Ryota Uehara; Akiko Tomioka; Chikashi Obuse; Minoru Kubo; Yuji Hiwatashi; Gohta Goshima
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-04-13
Journal Detail:
Title:  The Plant cell     Volume:  24     ISSN:  1532-298X     ISO Abbreviation:  Plant Cell     Publication Date:  2012 Apr 
Date Detail:
Created Date:  2012-05-30     Completed Date:  2012-09-24     Revised Date:  2013-06-26    
Medline Journal Info:
Nlm Unique ID:  9208688     Medline TA:  Plant Cell     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1478-93     Citation Subset:  IM    
Affiliation:
Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8602, Japan.
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MeSH Terms
Descriptor/Qualifier:
Anaphase
Bryopsida / cytology,  genetics*,  metabolism*
Gene Knockdown Techniques
Genes, Plant / genetics
Humans
Microtubules / metabolism*
Mitotic Spindle Apparatus / metabolism*
Phenotype
Plant Proteins / metabolism*
Plants, Genetically Modified
Protein Transport
RNA Interference*
Reproducibility of Results
Tubulin / metabolism
Chemical
Reg. No./Substance:
0/Plant Proteins; 0/Tubulin
Comments/Corrections

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