title: Live cell dynamics of promyelocytic leukemia nuclear bodies upon entry into and exit from mitosis.
abstract: Promyelocytic leukemia nuclear bodies (PML NBs) have been proposed to be involved in tumor suppression, viral defense, DNA repair, and/or transcriptional regulation. To study the dynamics of PML NBs during mitosis, we developed several U2OS cell lines stably coexpressing PML-enhanced cyan fluorescent protein with other individual marker proteins. Using three-dimensional time-lapse live cell imaging and four-dimensional particle tracking, we quantitatively demonstrated that PML NBs exhibit a high percentage of directed movement when cells progressed from prophase to prometaphase. The timing of this increased dynamic movement occurred just before or upon nuclear entry of cyclin B1, but before nuclear envelope breakdown. Our data suggest that entry into prophase leads to a loss of tethering between regions of chromatin and PML NBs, resulting in their increased dynamics. On exit from mitosis, Sp100 and Fas death domain-associated protein (Daxx) entered the daughter nuclei after a functional nuclear membrane was reformed. However, the recruitment of these proteins to PML NBs was delayed and correlated with the timing of de novo PML NB formation. Together, these results provide insight into the dynamic changes associated with PML NBs during mitosis.
authors.affiliation: Molecular and Cellular Biology Program, Stony Brook University, Stony Brook, NY 11794, USA.
authors.forename: Yi-Chun M
authors.initials: YC
authors.lastname: Chen
authors.forename: Constantin
authors.initials: C
authors.lastname: Kappel
authors.forename: Joel
authors.initials: J
authors.lastname: Beaudouin
authors.forename: Roland
authors.initials: R
authors.lastname: Eils
authors.forename: David L
authors.initials: DL
authors.lastname: Spector
mesh_terms: D034961:Antigens, Nuclear / Q000378:metabolism; D001324:Autoantigens / Q000378:metabolism; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000737:chemistry; D019926:Cyclin B / Q000378:metabolism; D056744:Cyclin B1; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D045586:Intranuclear Inclusion Bodies / Q000378:metabolism*; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism*; D008677:Metaphase; D008856:Microscopy, Fluorescence / Q000379:methods; D008938:Mitosis; D011418:Prophase; D013997:Time Factors; D014158:Transcription, Genetic
chemical_list: D034961:Antigens, Nuclear; D001324:Autoantigens; C534699:CCNB1 protein, human; D002843:Chromatin; C485182:Cyan Fluorescent Protein; D019926:Cyclin B; D056744:Cyclin B1; C066642:SP100 protein, human; D049452:Green Fluorescent Proteins