QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

Vol. 19, Issue 7, 3147-3162, July 2008

This Article Full Text Full Text (PDF) Supplemental Materials All Versions of this Article: E08-01-0035v1 19/7/3147    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Chen, Y.-C. M. Articles by Spector, D. L. PubMed PubMed Citation Articles by Chen, Y.-C. M. Articles by Spector, D. L.

Live Cell Dynamics of Promyelocytic Leukemia Nuclear Bodies upon Entry into and Exit from Mitosis

Yi-Chun M. Chen^*,, Constantin Kappel, Joel Beaudouin, Roland Eils^,, and David L. Spector^*,

*Molecular and Cellular Biology Program, Stony Brook University, Stony Brook, NY 11794; Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724; Division of Theoretical Bioinformatics, German Cancer Research Center, D-69120 Heidelberg, Germany; and Institute for Pharmacy and Molecular Biotechnology, University of Heidelberg, D-69120 Heidelberg, Germany

Submitted January 16, 2008; Revised April 22, 2008; Accepted May 1, 2008
Monitoring Editor: A. Gregory Matera

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.

Address correspondence to: David L. Spector (spector{at}cshl.edu)

Abbreviations used: CA, calyculin A; HCC, hypercondensed chromatin; IBB, importin-βbinding; MAPP, mitotic accumulations of PML protein; MLE, maximum-likelihood estimator; MSD, mean square displacement; NEBD, nuclear envelope breakdown; PMLNB, promyelocytic leukemia nuclear body.