Document Detail

Coupling of posterior cytoskeletal morphogenesis to the G1/S transition in the Trypanosoma brucei cell cycle.
MedLine Citation:
PMID:  15525678     Owner:  NLM     Status:  MEDLINE    
The expression levels of four Cdc2-related kinases (CRK1, 2, 4, and 6) in the procyclic form of Trypanosoma brucei were knocked down in pairs using the RNA interference (RNAi) technique. A double knockdown of CRK1 and CRK2 resulted in arrested cell growth in the G1 phase accompanied by an apparent cessation of nuclear DNA synthesis. The arrested cells became elongated at the posterior end like the G1-phase cells generated by knockdown of CycE1/CYC2 in a previous study. However, approximately 5% of the G1 cells in the current study also possessed multiply branched posterior ends, which have not previously been observed in T. brucei. DAPI and immunofluorescence staining showed a single nucleus, kinetoplast, basal body, and flagellum in the anterior portion of each G1 cell. The split and grossly extended posterior ends were heavily stained with antibodies to tyrosinated alpha-tubulin, suggesting an accumulation of newly synthesized microtubules. A significant population of anucleate cells (zoids), apparently derived from kinetoplast-dictated cytokinesis and cell division of the G1 cells, also had extended and branched posterior ends filled with newly synthesized microtubules. This continued posterior extension of microtubules in the G1 cells and zoids suggests that CRK1 and CRK2 exert a coordinated control on G1/S passage and the limited growth of the microtubule corset toward the posterior end. This connection may provide a new insight into the mechanism of morphological maintenance of an ancient protist during its cell cycle progression.
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.     Date:  2004-11-03
Journal Detail:
Title:  Molecular biology of the cell     Volume:  16     ISSN:  1059-1524     ISO Abbreviation:  Mol. Biol. Cell     Publication Date:  2005 Jan 
Date Detail:
Created Date:  2004-12-22     Completed Date:  2005-07-06     Revised Date:  2013-04-18    
Medline Journal Info:
Nlm Unique ID:  9201390     Medline TA:  Mol Biol Cell     Country:  United States    
Other Details:
Languages:  eng     Pagination:  97-105     Citation Subset:  IM    
Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94143-2280, USA.
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MeSH Terms
Bromodeoxyuridine / pharmacology
Cell Cycle
Cell Line
Cell Nucleus / metabolism
Cell Separation
Coloring Agents / pharmacology
Cytoskeleton / metabolism*
DNA / metabolism
DNA, Complementary / metabolism
DNA, Kinetoplast / metabolism
Disease Progression
Flagella / metabolism
Flow Cytometry
G1 Phase
Indoles / pharmacology
Microscopy, Fluorescence
RNA Interference
Reverse Transcriptase Polymerase Chain Reaction
S Phase
Time Factors
Trypanosoma brucei brucei
Tubulin / metabolism
Grant Support
Reg. No./Substance:
0/Coloring Agents; 0/DNA, Complementary; 0/DNA, Kinetoplast; 0/Indoles; 0/Tubulin; 47165-04-8/DAPI; 59-14-3/Bromodeoxyuridine; 9007-49-2/DNA

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