Trypanosoma brucei MOB1 is required for accurate and efficient cytokinesis but not for exit from mitosis.
Two MOB1 genes, MOB1-A and MOB1-B, were identified in Trypanosoma brucei. MOB1-A of T. brucei was shown to form a complex with TbPK50, a functional homologue of the Schizosaccharomyces pombe protein kinase Orb6, and immune precipitated MOB1-A exhibited histone H1 protein kinase activity. MOB1-A and TbPK50 were also shown to bind p12cks1, a cyclin-dependent kinase accessory protein. Immune fluorescence of epitope-tagged MOB1-A and MOB1-B in bloodstream form trypanosomes showed they had a punctate distribution all through the cell cytoplasm and were excluded from the nucleus throughout the cell cycle. Using RNA interference (RNAi), MOB1 was shown to be essential in both bloodstream and procyclic life cycle stages. In the bloodstream form, RNAi of MOB1 resulted, after 8 h, in a significant increase in post-mitotic cells, the majority of which had a visible cleavage furrow. This was followed by the appearance of cells with abnormal complements of nuclei and kinetoplasts, often with the number of nuclei exceeding the number of kinetoplasts. Thus, downregulation of MOB1 in the bloodstream form results in a delay in cytokinesis, and leads to a deregulation of the cell cycle, possibly through an inhibitory effect on kinetoplast replication. In contrast, downregulation of MOB1 in the procyclic form appears to impede the accuracy of cytokinesis, by allowing mispositioning of the cleavage furrow and inappropriate cytokinesis. Unlike its counterpart in budding yeast, T. brucei MOB1 does not appear to be required for mitotic exit.
Hammarton
Tansy C
TC
Wellcome Centre for Molecular Parasitology, The Anderson College, University of Glasgow, 56 Dumbarton Road, Glasgow G11 6NU, UK.
Lillico
Simon G
SG
Welburn
Sue C
SC
Mottram
Jeremy C
JC
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D048749:Cytokinesis / Q000502:physiology*; D005786:Gene Expression Regulation; D008018:Life Cycle Stages; D008938:Mitosis; D008969:Molecular Sequence Data; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*
C503177:MOB1-B protein, Trypanosoma brucei; D015800:Protozoan Proteins; D011494:Protein Kinases; C503176:MOB1-A protein, Trypanosoma brucei