Association of calmodulin with cytoskeletal structures at different stages of HeLa cell division, visualized by a calmodulin-EGFP fusion protein.
The fusion protein of calmodulin (CaM) with the enhanced green fluorescent protein EGFP has been expressed in a stably transfected HeLa cell line in order to visualise the localisation of calmodulin during the cell cycle on a continuous basis in live cells, and for immunofluorescence colocalisation with cytoskeletal structures. High-resolution images of CaM-EGFP in the mitotic apparatus show the characteristic strongly convoluted structure of the centrosome. CaM-EGFP also apparently associates with both polar and mitotic microtubules, and with a specific intracentrosomal structure. During cytokinesis, CaM-EGFP is also found decorating selected oriented filaments in close proximity to microtubules in the midbody region. In interphase cells, it is seen with filamentous and punctuate localisation at the nuclear envelope. The intensity and continuity of the CaM-EGFP images suggest that a significant fraction of the cellular calmodulin remains attached to cytoplasmic structures during the cell cycle.
Erent
M
M
Division of Physical Biochemistry, National Institute for Medical Research, London, United Kingdom.
Pagakis
S
S
Browne
J P
JP
Bayley
P
P
D002147:Calmodulin / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D003599:Cytoskeleton / Q000502:physiology* / Q000648:ultrastructure; D049452:Green Fluorescent Proteins; D006367:HeLa Cells / Q000166:cytology* / Q000502:physiology*; D006801:Humans; D008164:Luminescent Proteins; D018613:Microscopy, Confocal; D011993:Recombinant Fusion Proteins
D002147:Calmodulin; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins