Document Detail


Pairwise knockdowns of cdc2-related kinases (CRKs) in Trypanosoma brucei identified the CRKs for G1/S and G2/M transitions and demonstrated distinctive cytokinetic regulations between two developmental stages of the organism.
MedLine Citation:
PMID:  15821135     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Expression of the cdc2-related kinase 3 (CRK3) together with expression of CRK1, -2, -4, or -6, were knocked down in pairs in the procyclic and bloodstream forms of Trypanosoma brucei, using the RNA interference technique. Double knockdowns of CRK3 and CRK2, CRK4, or CRK6 exerted significant growth inhibition and enriched the cells in G2/M phase, whereas a CRK3 plus CRK1 (CRK3 + CRK1) knockdown arrested cells in both G1/S and G2/M transitions. Thus, CRK1 and CRK3 are apparently the kinases regulating the G1/S and G2/M checkpoint passages, respectively, whereas the other CRKs are probably playing only minor roles in cell cycle regulation. A CRK1 + CRK2 knockdown in the procyclic form was found to cause aberrant posterior cytoskeletal morphogenesis (X. M. Tu and C. C. Wang, Mol. Biol. Cell 16:97-105, 2005). A CRK3 + CRK2 knockdown, however, did not lead to such a change, suggesting that CRK2 depletion can lead to the abnormal morphogenesis only when procyclic-form cells are arrested in the G1 phase. The G2/M-arrested procyclic form produces up to 20% stumpy anucleated cells (zoids) in the population, suggesting that cytokinesis and cell division are not blocked by mitotic arrest but are apparently driven to completion by the kinetoplast cycle. In the bloodstream form, however, G2/M arrest resulted in little zoid formation but, instead, enriched a population of cells each containing multiple kinetoplasts, basal bodies, and flagella and an aggregate of multiple nuclei, indicating failure in entering cytokinesis. The two different cytokinetic regulations between two distinct stage-specific forms of the same organism may provide an interesting and useful model for further understanding the evolution of cytokinetic control among eukaryotes.
Authors:
Xiaoming Tu; Ching C Wang
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Eukaryotic cell     Volume:  4     ISSN:  1535-9778     ISO Abbreviation:  Eukaryotic Cell     Publication Date:  2005 Apr 
Date Detail:
Created Date:  2005-04-11     Completed Date:  2005-08-30     Revised Date:  2012-08-01    
Medline Journal Info:
Nlm Unique ID:  101130731     Medline TA:  Eukaryot Cell     Country:  United States    
Other Details:
Languages:  eng     Pagination:  755-64     Citation Subset:  IM    
Affiliation:
Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143-2280, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
CDC2 Protein Kinase / physiology*
Cell Cycle*
Cytokinesis
Protozoan Proteins / physiology*
RNA Interference
Time Factors
Transfection
Trypanosoma brucei brucei / enzymology*
Grant Support
ID/Acronym/Agency:
AI-21786/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Protozoan Proteins; EC 2.7.11.22/CDC2 Protein Kinase; EC 2.7.11.22/crk1 protein,Trypanosoma brucei; EC 2.7.11.22/crk2 protein, Trypanosoma brucei
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