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A Trypanosoma brucei protein required for maintenance of the flagellum attachment zone and flagellar pocket ER domains.

Lacomble S, Vaughan S, Deghelt M, Moreira-Leite FF, Gull K - Protist (2011)

Bottom Line: We also identified TbVAP, a T. brucei protein whose knockdown by RNAi in procyclic form cells leads to a dramatic reduction in the FAZ ER, and in the ER associated with the flagellar pocket.The localisation of tagged TbVAP and the phenotype of TbVAP RNAi in procyclic form trypanosomes are consistent with a function for TbVAP in the maintenance of sub-populations of the ER associated with the FAZ and the flagellar pocket.Nevertheless, depletion of TbVAP did not affect cell viability or cell cycle progression.

View Article: PubMed Central - PubMed

Affiliation: Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.

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The viability and cell cycle progression of T. brucei procyclic cells is not affected by TbVAP RNAi. A) Population growth curve of the TbVAP RNAi cells in the presence (black) or absence (grey) of doxycycline in the culture medium. Population growth was followed through a period of 72 h. B) Cell cycle analysis of TbVAP RNAi cells in the presence (black) or absence (grey) of doxycycline in the culture medium. The graph displays the % of cells in the classic cell cycle stages (1K1N, 2K1N and 2K2N) 24, 48 and 72 h after RNAi induction.
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fig0035: The viability and cell cycle progression of T. brucei procyclic cells is not affected by TbVAP RNAi. A) Population growth curve of the TbVAP RNAi cells in the presence (black) or absence (grey) of doxycycline in the culture medium. Population growth was followed through a period of 72 h. B) Cell cycle analysis of TbVAP RNAi cells in the presence (black) or absence (grey) of doxycycline in the culture medium. The graph displays the % of cells in the classic cell cycle stages (1K1N, 2K1N and 2K2N) 24, 48 and 72 h after RNAi induction.

Mentions: Despite the impact of TbVAP RNAi on the architecture of the ER at the FAZ and at the flagellar pocket, cells where TbVAP RNAi was induced remained viable. Population growth analysis revealed no difference between induced and non-induced cells in regular culture conditions, throughout a period of 72 h of induction (Fig. 5A). DAPI-stained cells from an asynchronously dividing population can be placed into one of three stages of the cell cycle: 1) 1K1N cells, where neither the kinetoplast (K) nor the nucleus (N) have divided (corresponding to the G1/S stage of the cell cycle); 2) 2K1N cells, where kinetoplast (but not nuclear) division has occurred (G2/M phase cells); and 2K2N cells, where both the kinetoplast and the nucleus have divided, prior to cytokinesis (Sherwin and Gull 1989; Woodward and Gull 1990). Induction of TbVAP RNAi did not affect the proportion of cell cycle stages (Fig. 5B).


A Trypanosoma brucei protein required for maintenance of the flagellum attachment zone and flagellar pocket ER domains.

Lacomble S, Vaughan S, Deghelt M, Moreira-Leite FF, Gull K - Protist (2011)

The viability and cell cycle progression of T. brucei procyclic cells is not affected by TbVAP RNAi. A) Population growth curve of the TbVAP RNAi cells in the presence (black) or absence (grey) of doxycycline in the culture medium. Population growth was followed through a period of 72 h. B) Cell cycle analysis of TbVAP RNAi cells in the presence (black) or absence (grey) of doxycycline in the culture medium. The graph displays the % of cells in the classic cell cycle stages (1K1N, 2K1N and 2K2N) 24, 48 and 72 h after RNAi induction.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3405529&req=5

fig0035: The viability and cell cycle progression of T. brucei procyclic cells is not affected by TbVAP RNAi. A) Population growth curve of the TbVAP RNAi cells in the presence (black) or absence (grey) of doxycycline in the culture medium. Population growth was followed through a period of 72 h. B) Cell cycle analysis of TbVAP RNAi cells in the presence (black) or absence (grey) of doxycycline in the culture medium. The graph displays the % of cells in the classic cell cycle stages (1K1N, 2K1N and 2K2N) 24, 48 and 72 h after RNAi induction.
Mentions: Despite the impact of TbVAP RNAi on the architecture of the ER at the FAZ and at the flagellar pocket, cells where TbVAP RNAi was induced remained viable. Population growth analysis revealed no difference between induced and non-induced cells in regular culture conditions, throughout a period of 72 h of induction (Fig. 5A). DAPI-stained cells from an asynchronously dividing population can be placed into one of three stages of the cell cycle: 1) 1K1N cells, where neither the kinetoplast (K) nor the nucleus (N) have divided (corresponding to the G1/S stage of the cell cycle); 2) 2K1N cells, where kinetoplast (but not nuclear) division has occurred (G2/M phase cells); and 2K2N cells, where both the kinetoplast and the nucleus have divided, prior to cytokinesis (Sherwin and Gull 1989; Woodward and Gull 1990). Induction of TbVAP RNAi did not affect the proportion of cell cycle stages (Fig. 5B).

Bottom Line: We also identified TbVAP, a T. brucei protein whose knockdown by RNAi in procyclic form cells leads to a dramatic reduction in the FAZ ER, and in the ER associated with the flagellar pocket.The localisation of tagged TbVAP and the phenotype of TbVAP RNAi in procyclic form trypanosomes are consistent with a function for TbVAP in the maintenance of sub-populations of the ER associated with the FAZ and the flagellar pocket.Nevertheless, depletion of TbVAP did not affect cell viability or cell cycle progression.

View Article: PubMed Central - PubMed

Affiliation: Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.

Show MeSH
Related in: MedlinePlus