Abstract
The parasitic protozoan Trypanosoma brucei contains two type III phosphatidylinositol 4-kinases (α and β). We have cloned the gene encoding the T. brucei type III phosphatidylinositol 4-kinase β (TbPI4KIII-β), expressed the protein in COS-7 cells, and confirmed that the protein catalyzes the phosphorylation of phosphatidylinositol. Depletion of TbPI4KIII-β in procyclic T. brucei by RNA interference (RNAi) resulted in inhibition of cell growth and a distorted cellular morphology. RNAi cells had a distorted Golgi apparatus, and lysosomal and flagellar pocket proteins were mislocalized. Ultrastructural analysis revealed the internal accumulation of a heterogeneous population of vesicles, abnormal positioning of organelles, and a loss of cell polarity. Scanning electron microcopy revealed a twisted phenotype, and dividing cells often exhibited a detached daughter flagellum and lacked a cleavage furrow. Cell cycle analysis confirmed that cells depleted of TbPI4KIII-β have a postmitotic cytokinesis block that occurs after a single round of mitosis, suggestive of a specific cell cycle block. In summary, TbPI4KIII-β is an essential protein in procyclic T. brucei, required for maintenance of Golgi structure, protein trafficking, normal cellular shape, and cytokinesis.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1108-1118 |
| Number of pages | 11 |
| Journal | Eukaryotic Cell |
| Volume | 6 |
| Issue number | 7 |
| DOIs | |
| State | Published - Jul 2007 |
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ASJC Scopus subject areas
- Agricultural and Biological Sciences(all)
- Microbiology
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Phosphatidylinositol 4-kinase III-β is required for Golgi maintenance and cytokinesis in Trypanosoma brucei. / Rodgers, Melissa J.; Albanesi, Joseph P.; Phillips, Margaret A.
In: Eukaryotic Cell, Vol. 6, No. 7, 07.2007, p. 1108-1118.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Phosphatidylinositol 4-kinase III-β is required for Golgi maintenance and cytokinesis in Trypanosoma brucei
AU - Rodgers, Melissa J.
AU - Albanesi, Joseph P.
AU - Phillips, Margaret A.
PY - 2007/7
Y1 - 2007/7
N2 - The parasitic protozoan Trypanosoma brucei contains two type III phosphatidylinositol 4-kinases (α and β). We have cloned the gene encoding the T. brucei type III phosphatidylinositol 4-kinase β (TbPI4KIII-β), expressed the protein in COS-7 cells, and confirmed that the protein catalyzes the phosphorylation of phosphatidylinositol. Depletion of TbPI4KIII-β in procyclic T. brucei by RNA interference (RNAi) resulted in inhibition of cell growth and a distorted cellular morphology. RNAi cells had a distorted Golgi apparatus, and lysosomal and flagellar pocket proteins were mislocalized. Ultrastructural analysis revealed the internal accumulation of a heterogeneous population of vesicles, abnormal positioning of organelles, and a loss of cell polarity. Scanning electron microcopy revealed a twisted phenotype, and dividing cells often exhibited a detached daughter flagellum and lacked a cleavage furrow. Cell cycle analysis confirmed that cells depleted of TbPI4KIII-β have a postmitotic cytokinesis block that occurs after a single round of mitosis, suggestive of a specific cell cycle block. In summary, TbPI4KIII-β is an essential protein in procyclic T. brucei, required for maintenance of Golgi structure, protein trafficking, normal cellular shape, and cytokinesis.
AB - The parasitic protozoan Trypanosoma brucei contains two type III phosphatidylinositol 4-kinases (α and β). We have cloned the gene encoding the T. brucei type III phosphatidylinositol 4-kinase β (TbPI4KIII-β), expressed the protein in COS-7 cells, and confirmed that the protein catalyzes the phosphorylation of phosphatidylinositol. Depletion of TbPI4KIII-β in procyclic T. brucei by RNA interference (RNAi) resulted in inhibition of cell growth and a distorted cellular morphology. RNAi cells had a distorted Golgi apparatus, and lysosomal and flagellar pocket proteins were mislocalized. Ultrastructural analysis revealed the internal accumulation of a heterogeneous population of vesicles, abnormal positioning of organelles, and a loss of cell polarity. Scanning electron microcopy revealed a twisted phenotype, and dividing cells often exhibited a detached daughter flagellum and lacked a cleavage furrow. Cell cycle analysis confirmed that cells depleted of TbPI4KIII-β have a postmitotic cytokinesis block that occurs after a single round of mitosis, suggestive of a specific cell cycle block. In summary, TbPI4KIII-β is an essential protein in procyclic T. brucei, required for maintenance of Golgi structure, protein trafficking, normal cellular shape, and cytokinesis.
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U2 - 10.1128/EC.00107-07
DO - 10.1128/EC.00107-07
M3 - Article
VL - 6
SP - 1108
EP - 1118
JO - Eukaryotic Cell
JF - Eukaryotic Cell
SN - 1535-9778
IS - 7
ER -