Centrin gene disruption impairs stage-specific basal body duplication and cell cycle progression in Leishmania

Angamuthu Selvapandiyan, Alain Debrabant, Robert Duncan, Jacqueline Muller, Poonam Salotra, Gannavaram Sreenivas, Jeffrey L Salisbury, Hira L. Nakhasi

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

Centrin is a calcium-binding cytoskeletal protein involved in the duplication of centrosomes in higher eukaryotes. To explore the role of centrin in the protozoan parasite Leishmania, we created Leishmania deficient in the centrin gene (LdCEN). Remarkably, centrin null mutants (LdCEN -/-) showed selective growth arrest as axenic amastigotes but not as promastigotes. Flow cytometry analysis confirmed that the mutant axenic amastigotes have a cell cycle arrest at the G 2/M stage. The axenic amastigates also showed failure of basal body duplication and failure of cytokinesis resulting in multinucleated "large" cells. Increased terminal deoxy uridine triphosphate nick end labeling positivity was observed in centrin mutant axenic amastigotes compared with wild type cells, suggesting the activation of a programmed cell death pathway. Growth of LdCEN -/- amastigotes in infected macrophages in vitro was inhibited and also resulted in large multinucleated parasites. Normal basal body duplication and cell division in the LdCEN knockout promastigote is unique and surprising. Further, this is the first report where disruption of a centrin gene displays stage-specific/cell type-specific failure in cell division in a eukaryote. The centrin null mutant defective in amastigote growth could be useful as a vaccine candidate against leishmaniasis.

Original languageEnglish (US)
Pages (from-to)25703-25710
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number24
DOIs
StatePublished - Jun 11 2004

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Basal Bodies
Leishmania
Sulfamethoxazole Drug Combination Trimethoprim
Cell Cycle
Genes
Cells
Eukaryota
Cell Division
Parasites
Growth
Centrosome
Gene Knockout Techniques
Uridine Triphosphate
Calcium-Binding Proteins
Leishmaniasis
Cytoskeletal Proteins
Cytokinesis
Flow cytometry
Macrophages
Cell death

ASJC Scopus subject areas

  • Biochemistry

Cite this

Selvapandiyan, A., Debrabant, A., Duncan, R., Muller, J., Salotra, P., Sreenivas, G., ... Nakhasi, H. L. (2004). Centrin gene disruption impairs stage-specific basal body duplication and cell cycle progression in Leishmania. Journal of Biological Chemistry, 279(24), 25703-25710. https://doi.org/10.1074/jbc.M402794200

Centrin gene disruption impairs stage-specific basal body duplication and cell cycle progression in Leishmania. / Selvapandiyan, Angamuthu; Debrabant, Alain; Duncan, Robert; Muller, Jacqueline; Salotra, Poonam; Sreenivas, Gannavaram; Salisbury, Jeffrey L; Nakhasi, Hira L.

In: Journal of Biological Chemistry, Vol. 279, No. 24, 11.06.2004, p. 25703-25710.

Research output: Contribution to journalArticle

Selvapandiyan, A, Debrabant, A, Duncan, R, Muller, J, Salotra, P, Sreenivas, G, Salisbury, JL & Nakhasi, HL 2004, 'Centrin gene disruption impairs stage-specific basal body duplication and cell cycle progression in Leishmania', Journal of Biological Chemistry, vol. 279, no. 24, pp. 25703-25710. https://doi.org/10.1074/jbc.M402794200
Selvapandiyan, Angamuthu ; Debrabant, Alain ; Duncan, Robert ; Muller, Jacqueline ; Salotra, Poonam ; Sreenivas, Gannavaram ; Salisbury, Jeffrey L ; Nakhasi, Hira L. / Centrin gene disruption impairs stage-specific basal body duplication and cell cycle progression in Leishmania. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 24. pp. 25703-25710.
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