The Trypanosoma brucei cyclin, CYC2, is required for cell cycle progression through G1 phase and for maintenance of procyclic form cell morphology.
CYC2 is an essential PHO80-like cyclin that forms a complex with the cdc2-related kinase CRK3 in Trypanosoma brucei. In both procyclic and bloodstream form T. brucei, knock-down of CYC2 by RNA interference (RNAi) led to an accumulation of cells in G(1) phase. Additionally, in procyclic cells, but not in bloodstream form cells, CYC2 RNAi induced a specific cell elongation at the posterior end. The G(1) block, as well as the posterior end elongation in the procyclic form, was irreversible once established. Staining for tyrosinated alpha-tubulin and morphometric analyses showed that the posterior end elongation occurred through active microtubule extension, with no repositioning of the kinetoplast. Hence, these cells can be classified as exhibiting the "nozzle" phenotype as has been described for cells that ectopically express TbZFP2, a zinc finger protein that is involved in the differentiation of the bloodstream form to procyclic form. Within the tsetse fly, procyclic trypanosomes differentiate to elongated mesocyclic cells. However, although mesocyclic trypanosomes isolated from tsetse flies also show active microtubule extension at the posterior end, the kinetoplast is coincidentally repositioned such that it always lies approximately midway between the nucleus and posterior end of the cell. Thus, in the procyclic form CYC2 has dual functionality and is required for both cell cycle progression through G(1) and for the maintenance of correct cell morphology, whereas in the bloodstream form only a role for CYC2 in G(1) progression is evident.
Hammarton
Tansy C
TC
Wellcome Centre for Molecular Parasitology, the Anderson College, University of Glasgow, 56 Dumbarton Road, Glasgow G11 6NU, Scotland, United Kingdom.
Engstler
Markus
M
Mottram
Jeremy C
JC
D000818:Animals; D002453:Cell Cycle; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D016213:Cyclins / Q000378:metabolism / Q000502:physiology*; D004247:DNA / Q000378:metabolism; D018105:DNA, Kinetoplast; D018450:Disease Progression; D005434:Flow Cytometry; D016193:G1 Phase; D015151:Immunoblotting; D008870:Microtubules / Q000378:metabolism; D010641:Phenotype; D015800:Protozoan Proteins / Q000378:metabolism / Q000502:physiology*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D014404:Tubulin / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry; D016335:Zinc Fingers
D016213:Cyclins; D018105:DNA, Kinetoplast; D015800:Protozoan Proteins; D012333:RNA, Messenger; D014404:Tubulin; C402347:cyc2 protein, Paramecium tetraurelia; D014443:Tyrosine; D004247:DNA